Sample records for culex pipiens evokes

Background Culex spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, Culexpipienspipiens, Culexpipiens quinquefasciatus, and Culex tarsalis are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of Culexpipiens complex mosquitoes and hybrids in Mexico City. Culexpipienspipiens preferentially feed on avian hosts; Culexpipiens quinquefasciatus have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of Culexpipiens complex mosquitoes and their hybrids. Results Mosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as Culex, Culiseta or Ochlerotatus by morphological analysis. Within the genus Culex, specimens were further identified as Culex tarsalis or as belonging to the Culexpipiens complex. Members of the Culexpipiens complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the Ace.2 gene. Culexpipiens quinquefasciatus was the most abundant form collected. Conclusions Important WNV vectors species, Cx. p. pipiens, Cx. p. quinquefasciatus and Cx. tarsalis, are all present in Mexico City. Hybrids of Cx. p. pipiens and Cx. p. quinquefasciatus were also

Overwintering populations of Culexpipiens, the principal enzootic vector of West Nile virus in the northeastern USA, were studied over 3 consecutive winters from 2006 to 2008, using mark-recapture techniques to determine when Cx. pipiens females began to disperse from overwintering hibernacula and how their survival influenced early season populations. In February of each year, Cx. pipiens were aspirated and marked using fluorescent powder; 4,067, 752, and 3,070 diapausing Cx. pipiens were marked in each successive year. Mosquitoes were then trapped from mid-April to early May of each year using 19 Centers for Disease Control and Prevention (CDC) light traps and 16 CDC gravid traps. A total of 348, 39, and 111 Culex mosquitoes were captured in the spring of 2006, 2007, and 2008, respectively. The number of mosquitoes marked in overwintering habitats is generally positively correlated with the number of mosquitoes recaptured in the early spring (linear regression, R2 = 0.79, P = 0.04), yet results also suggest that seasonal variations beyond overwintering population size are likely important in determining the success of emergent populations. A single marked Cx. pipiens was captured in both 2006 and 2008. In 2006, the mosquito was captured 0.5 km from its overwintering site while in 2008 the mosquito was captured 0.3 km from its overwintering site. In all study years, mosquitoes consistently began exiting overwintering hibernacula the 3rd week of April, yet evidence of earlier exodus was observed in 2007, when outside temperatures were significantly higher in preceding days and months.

Mosquitoes in the Culexpipiens complex thrive in temperate and tropical regions worldwide, and serve as efficient vectors of Bancroftian lymphatic filariasis (LF) caused by Wuchereria bancrofti in Asia, Africa, the West Indies, South America, and Micronesia. However, members of this mosquito complex do not act as natural vectors for Brugian LF caused by Brugia malayi, or for the cat parasite B. pahangi, despite their presence in South Asia where these parasites are endemic. Previous work with the Iowa strain of Culexpipienspipiens demonstrates that it is equally susceptible to W. bancrofti as is the natural Cx. p. pipiens vector in the Nile Delta, however it is refractory to infection with Brugia spp. Here we report that the infectivity barrier for Brugia spp. in Cx. p. pipiens is the mosquito midgut, which inflicts internal and lethal damage to ingested microfilariae. Following per os Brugia exposures, the prevalence of infection is significantly lower in Cx. p. pipiens compared to susceptible mosquito controls, and differs between parasite species with <50% and <5% of Cx. p. pipiens becoming infected with B. pahangi and B. malayi, respectively. When Brugia spp. mf were inoculated intrathoracically to bypass the midgut, larvae developed equally well as in controls, indicating that, beyond the midgut, Cx. p. pipiens is physiologically compatible with Brugia spp. Mf isolated from Cx. p. pipiens midguts exhibited compromised motility, and unlike mf derived from blood or isolated from the midguts of Ae. aegypti, failed to develop when inoculated intrathoracically into susceptible mosquitoes. Together these data strongly support the role of the midgut as the primary infection barrier for Brugia spp. in Cx. p. pipiens. Examination of parasites recovered from the Cx. p. pipiens midgut by vital staining, and those exsheathed with papain, suggest that the damage inflicted by the midgut is subcuticular and disrupts internal tissues. Microscopic studies of these worms

The common house mosquito, Culexpipiens (Diptera: Culicidae), which is considered the primary bridge vector of West Nile Virus (WNV) to humans, is a wide spread insect pest with medical importance and consists of two distinct bioforms, Cx. pipiens f. pipiens and Cx. pipiens f. molestus. Here, we consider the adult lifespan of male and female Cx. pipiens f. pipiens under controlled conditions at five constant temperature regimes (15, 20, 25, 27.5, and 30 °C). Our results show that adult longevity was affected by temperature, as it significantly decreased with increase in temperature. At the highest tested temperature, mean adult longevity did not exceed 12 days for both sexes and thus makes impossible the risk of WNV transmission. On the other hand at the lowest temperature, longevity was extremely high with some individuals surviving up to 129 and 132 days, males and females, respectively, and thus enable them to function as potential vectors of WNV for a prolonged period of time. As far as sex is concerned, adult females displayed a 1.2-1.4-fold longer longevity compared to the male ones. However, this difference was significant only at the lowest and highest tested temperature regime. This information is useful in determining the critical temperatures which may affect the distribution of Cx. pipiens and consequently the risk of WNV transmission. Moreover, the effect of environmental temperature should be considered when evaluating the abundance of these species.

The Culexpipiens complex, including Culexpipiens, Cx. quinquefasciatus, and Cx. molestus are important pest species and vectors of human and animal diseases throughout the world's tropical, temperate, and Holarctic regions. Diseases transmitted by member of the complex include: St. Louis encephali...

Mosquito larvae face numerous biotic and abiotic challenges that affect their development and survivorship, as well as adult fitness. We conducted two experiments under semi-natural conditions to evaluate the effects of intraspecific competition, nutrient limitation and sub-lethal doses of malathion on individual life history traits in adult Culexpipiens (Diptera: Culicidae). In the first experiment, larvae of Cx. pipiens were reared at different intraspecific densities and exposed to sub-lethal doses of malathion. In the second experiment, different intraspecific densities of Cx. pipiens larvae were reared under conditions of low or high larval nutrients, and subsequent adults were fed on either water or 10% sucrose solution. Malathion treatment had relatively minor effects compared with density, which had significant negative effects on development rate, survivorship to adulthood, body size (wing length) and longevity. As larval density increased, a sex ratio distortion in survivorship to adulthood emerged, in which a bias towards males was apparent. Nutrient-rich larval environments alleviated, in part, the effects of increasing density and extended the lifespan of mosquitoes fed on water and 10% sucrose. Density-dependent alterations in adult longevity attributable to the larval environment are complex and show contrasting results depending on interactions with other environmental factors. This study suggests that larval resource availability and competition influence Cx. pipiens population growth correlates and have lasting effects on traits that relate to a mosquito's ability to vector pathogens.

The climatic and housing conditions which favour the breeding of Culexpipiens fatigans in Ceylon, and its life-cycle and resting-habits, are described. The results of the treatment of breeding-places (mainly catch-pits) and the interior of houses with various insecticides of specific action on larvae or adults, in different preparations and concentrations, are given, and suggestions are made for the control of the mosquito on this island. The fundamental measure for successful control is efficient sanitation, but until this can be achieved, application of larvicides seems to be the method of choice. PMID:13472415

Females of the northern house mosquito, Culexpipiens L., are capable of entering an adult overwintering diapause characterized by arrested ovarian development, enhanced stress tolerance, and elevated lipid stores. In contrast, the southern house mosquito, Culex quinquefasciatus Say, lacks this capacity and is therefore unable to survive the harsh winters found in northern regions of North America. These two species are capable of forming fertile hybrids in the United States, yet the diapause characteristics of these hybrids have not been extensively investigated. We crossed Cx. pipiens from Columbus, OH, with Cx. quinquefasciatus from Vero Beach, FL, and reared F1 hybrids from all mothers separately under diapause-inducing, short-day conditions (a photoperiod of 8:16 [L:D] h) at 18°C. Egg follicle length and lipid content were used to assess the diapause status of hybrids. Diapause incidence of hybrids varied widely for progeny from different mothers of the same species, but hybrids with Cx. pipiens mothers were consistently more prone to enter diapause than hybrids that had Cx. quinquefasciatus mothers. Our results suggest a strong maternal influence on the diapause phenotype and that a high percentage (45-75%) of Cx. pipiens-Cx. quinquefasciatus hybrids are capable of entering diapause. This implies that many hybrids can successfully overwinter, leading to a possible widening of the hybrid zone of these two species in North America.

Microsatellite markers were used to genetically characterize 19 Culexpipiens complex populations from California. Two populations showed characteristics of earlier genetic bottlenecks. The overall FST value and a neighbor-joining tree suggested moderate amounts of genetic differentiation. Analyses using Structure indicated K = 4 genetic clusters: Cx. pipiens form pipiens L., Cx. quinquefasciatus Say, Cx. pipiens form molestus Forskäl, and a group of genetically similar individuals of hybrid origin. A Discriminant Analysis of Principal Components indicated that the latter group is a mixture of the other three taxa, with form pipiens and form molestus contributing somewhat more ancestry than Cx. quinquefasciatus. Characterization of 56 morphologically autogenous individuals classified most as Cx. pipiens form molestus, and none as Cx. pipiens form pipiens or Cx. quinquefasciatus. Comparison of California microsatellite data with those of Cx. pipiens pallens Coquillett from Japan indicated the latter does not contribute significantly to genotypes in California. PMID:23958909

Background: The role of lectins is important in interaction between pathogens and mosquito vectors. This study was performed to identify agglutinin activities of protein molecules on the midgut of Culexpipiens. Methods: Culexpipiens was reared in insectray condition and the midguts of males and females (blood fed and unfed) were dissected separately in Tris-HCl buffer. The extracts of midguts were applied for hemagglutinin assay against red blood cells of rabbit, mouse, rat, dog, horse, sheep, guinea pig, cow, human (A, B, AB, O groups). Then, the RBCs with relatively high agglutinin activity were chosen for carbohydrate inhibition assay. D (+) glucose, D (+) galactose, D (+) mannose, D (−) fructose, D (−) arabinose, L (−) fucose, lactose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, sialic acid were used to specify carbohydrate binding lectin. Results: The highest agglutinin activities were found against sheep and rabbits RBCs. Sexual diversity of agglutinin activities was observed among midgut extraction of males and females. In addition, variation in agglutinin activity of blood fed and unfed female mosquitoes were detected. The lectin activity was inhibited highly with glucose, galactose, fucose and fructose but less inhibitor activities was observed by arabinose, N-acetyl-D-galactosamine, n-acetyl-d-glucosamine, lactose and mannose. Conclusion: The secretion of hemagglutinins (lectins or lectin-like molecules) in the digestive system depends on the type of food in the gut. This suggests that emptying of the gut in preparation for protein rich food probably starts the secretion of hemagglutinins. PMID:23785692

Abstract Nucleotide sequences of the circadian rhythm genes, period and timeless, were studied for the first time in mosquitoes Culexpipiens Linnaeus, 1758. In this work we evaluated variations of the studied genome fragments for the two forms of Culexpipiens (forma “pipiens” – mosquitoes common for aboveground habitats, forma “molestus” – underground mosquitoes). We compared Culexpipiens from Russia with transatlantic Culexpipiens and subtropical Culex quinquefasciatus Say, 1823. Our results show that intraspecies variability is higher for the gene period than for the gene timeless. The revealed substitutions in nucleotide sequences and especially in amino acid sequences grouped the individuals of the two forms into distinct clusters with high significance. The detected fixed amino acid substitutions may appear essential for functioning of the circadian rhythm proteins in Culexpipiens, and may be correlated with adaptations of the taxa within the group Culexpipiens. Our results suggest that natural selection favors fixed mutations and the decrease in diversity of the genes period and timeless in mosquitoes of the Culexpipiens f. “molestus” compared with the Culexpipiens f. “pipiens”, is probably correlated with adaptive features of Culexpipiens f. “molestus”. The studied genome regions may be considered as promising molecular-genetic markers for identification, population and phylogenetic analysis of similar species and forms of the Culexpipiens complex. PMID:28123673

Zika virus (ZIKV) has become a threat to global health since the outbreak in Brazil in 2015. Although ZIKV is generally considered an Aedes-transmitted pathogen, new evidence has shown that parts of the virus closely resemble Culex-transmitted viruses. Therefore, it is important to evaluate the competence of Culex species for ZIKV to understand their potential as vectors. In this study, female Culexpipiens quinquefasciatus were orally exposed to ZIKV. Mosquito midguts, salivary glands and ovaries were tested for ZIKV to measure infection and dissemination at 2, 4, 6, 8, 12, 16 and 18 days post exposure (pe). In addition, saliva was collected from mosquitoes after infection and infant mice were bitten by infected mosquitoes to measure the transmission ability of Cx. p. quinquefasciatus. The results showed that the peak time of virus appearance in the salivary glands was day 8 pe, with 90% infection rate and an estimated virus titer of 3.92±0.49 lg RNA copies/mL. Eight of the nine infant mice had positive brains after being bitten by infected mosquitoes, which meant that Cx. p. quinquefasciatus could be infected with and transmit ZIKV following oral infection. These laboratory results clearly demonstrate the potential role of Cx. p. quinquefasciatus as a vector of ZIKV in China. Because there are quite different vector management strategies required to control Aedes (Stegomyia) species and Cx. p. quinquefasciatus, an integrated approach may be required should a Zika epidemic occur.

As a result of evidence of the ecotoxicity of nitrofurans, the acute toxicity of furazolidone was tested in vivo on two aquatic organisms, Artemia salina and Daphnia magna, which are both crustaceans. Toxicity studies were also performed on larvae of Culexpipiens molestus. Results indicated a significant toxicity of the compound on Culexpipiens and Daphnia magna, while Artemia salina proved to be the least sensitive.

Observations have been made on Culexpipiens fatigans in the Taveta-Pare area in East Africa from 1954 to 1961, during which time dieldrin was applied to local houses over a 3½-year period as part of an experiment in malaria control. At the end of the period of residual spraying the numbers of C. p. fatigans had been reduced by two-thirds in the Taveta area, but in the South Pare area, only some 160 km away, this mosquito was twice as numerous as before spraying. The results of susceptibility tests carried out in untreated and dieldrin-treated areas showed that the susceptibility of C. p. fatigans in East Africa to chlorinated hydrocarbon insecticides was low compared with that of Anopheles gambiae, and varied by at least 20 times in some areas relatively short distances apart. C. p. fatigans from all areas sampled were, however, susceptible to fenthion and malathion. PMID:13993106

We investigated the susceptibility of an Italian population of Culexpipiens mosquitoes to Zika virus (ZIKV) infection, tested in parallel with Aedes aegypti, as a positive control. We analysed mosquitoes at 0, 3, 7, 10, 14, 20 and 24 days after an infectious blood meal. Viral RNA was detected in the body of Cx. pipiens up to three days post-infection, but not at later time points. Our results indicate that Cx. pipiens is not susceptible to ZIKV infection. PMID:27605056

We investigated the susceptibility of an Italian population of Culexpipiens mosquitoes to Zika virus (ZIKV) infection, tested in parallel with Aedes aegypti, as a positive control. We analysed mosquitoes at 0, 3, 7, 10, 14, 20 and 24 days after an infectious blood meal. Viral RNA was detected in the body of Cx. pipiens up to three days post-infection, but not at later time points. Our results indicate that Cx. pipiens is not susceptible to ZIKV infection.

Background: Culexpipiens molestus was first reported in Shanghai in 2010. The population structures and seasonal distributions of Culexpipiens subspecies C. p. molestus, Culexpipiens pallens, and Culexpipiens quinquefasciatus are not well known. Methods: From late February to November 2013, we conducted daily field surveillance of mosquitoes at eight sites at two green lands and three residential areas in downtown Shanghai. Morphological comparison and DV/D ratios (DV/D is an indicator of mosquito taxonomy) were used to identify adult mosquitoes. Results: The distribution curves of the Culexpipiens complex members indicated seasonal fluctuations. The temperature range of 20–25 °C was the most suitable for adult activity. Micro-environmental factors may differentiate the complex population structures. Hybridization between C. p. pallens and C. p. quinquefasciatus was common and neither “DV/D = 0.40” nor “DV/D = 0.50” can distinguish these subspecies and their hybrids. Conclusion: the population structure of the Culexpipiens complex is complex and characterized by significant hybridization. Measures other than DV/D ratios are needed for the discrimination of subspecies. The C. p. molestus invasion might result in the transmission of novel vector-borne diseases in Shanghai. PMID:27869687

The α-proteobacteria Wolbachia are among the most common intracellular bacteria and have recently emerged as important drivers of arthropod biology. Wolbachia commonly act as reproductive parasites in arthropods by inducing cytoplasmic incompatibility (CI), a type of conditional sterility between hosts harboring incompatible infections. In this study, we examined the evolutionary histories of Wolbachia infections, known as wPip, in the common house mosquito Culexpipiens, which exhibits the greatest variation in CI crossing patterns observed in any insect. We first investigated a panel of 20 wPip strains for their genetic diversity through a multilocus scheme combining 13 Wolbachia genes. Because Wolbachia depend primarily on maternal transmission for spreading within arthropod populations, we also studied the variability in the coinherited Cx. pipiens mitochondria. In total, we identified 14 wPip haplotypes, which all share a monophyletic origin and clearly cluster into five distinct wPip groups. The diversity of Cx. pipiens mitochondria was extremely reduced, which is likely a consequence of cytoplasmic hitchhiking driven by a unique and recent Wolbachia invasion. Phylogenetic evidence indicates that wPip infections and mitochondrial DNA have codiverged through stable cotransmission within the cytoplasm and shows that a rapid diversification of wPip has occurred. The observed pattern demonstrates that a considerable degree of Wolbachia diversity can evolve within a single host species over short evolutionary periods. In addition, multiple signatures of recombination were found in most wPip genomic regions, leading us to conclude that the mosaic nature of wPip genomes may play a key role in their evolution.

Background Blood feeding by free-living insect vectors of disease is rhythmic and can be used to predict when infectious bites will occur. These daily rhythms can also be targeted by control measures, as in insecticide-treated nets. Culexpipiens form pipiens and C.p. f. molestus are two members of the Culexpipiens assemblage and vectors of West Nile Virus throughout North America. Although Culex species vector human pathogens and parasites, the daily blood feeding rhythms of C.p. f. molestus, to our knowledge, have not been studied. We described and compared the daily blood feeding rhythms of three laboratory-reared populations of Culexpipiens, one of which has confirmed molestus ancestry. We also examined the plasticity of blood feeding time for these three populations. Results For most (>70%) C.p. f. pipiens and C.p. f. molestus collected from metropolitan Chicago, IL, blood feeding took place during scotophase. Peak blood feeding occurred in mid-scotophase, 3-6 hours after lights off. For C.p. f. pipiens originating from Pennsylvania, most mosquitoes (> 90%) blood fed during late photophase and early scotophase. C.p. f. molestus denied a blood meal during scotophase were less likely to blood feed during early photophase (< 20%) than were C.p. f. pipiens from Chicago (> 50%). C.p. f. pipiens from Pennsylvania were capable of feeding readily at any hour of photo- or scotophase. Conclusions Daily blood feeding rhythms of C.p. f. molestus are similar to those of C.p. f. pipiens, particularly when populations originate from the same geographic region. However, the timing of blood feeding is more flexible for C.p. f. pipiens populations relative to C.p. f. molestus. PMID:24450879

Ethanolic, acetone and petroleum ether extracts from leaves and stems of Lagenaria siceraria (Cucurbitaceae) were screened for their repellency effect against Culexpipiens L. mosquito. The repellent action of the present plant extracts were varied depending on the plant parts and the dose of extract. The petroleum ether extract of leaves showed the same repellency percent (100%) of commercial formulation, N. N.diethyl toulamide (DEET) at the higher dose (3.33 mg/cm2), while petroleum ether extract from stems exhibiting the repellent action (89.6%) at the same dose, respectively. Ethanolic extracts of leaves and stems exhibited the lowest repellent activity as it recorded (81.3% and 69.1%) at (6.67 mg/cm2), respectively. Results of this study may contribute to design an alternative way to control mosquitoes currently based on applications of synthetic insecticides. These extracts could be developed commercially as an effective personal protection measure against mosquito bites and thus to control diseases caused by mosquito-borne pathogens.

Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI), resulting in abortive embryonic development. One Wolbachia host, Culexpipiens complex mosquitoes, displays high levels of variability in both CI crossing types (cytotypes) and DNA markers. We report here an analysis of 14 mosquito strains, containing 13 Wolbachia variants, and with 13 different cytotypes. Cytotypes were Wolbachia-dependent, as antibiotic treatment rendered all strains tested compatible. Cytotype distributions were independent of geographical distance between sampling sites and host subspecies, suggesting that Wolbachia does not promote a reproductive isolation depending on these parameters. Backcross analysis demonstrated a mild restoring effect of the nuclear genome, indicating that CI is mostly cytoplasmically determined for some crosses. No correlation was found between the phenotypic and genotypic variability of 16 WO prophage and transposon markers, except for the WO prophage Gp15 gene, which encodes a protein similar to a bacterial virulence factor. However, Gp15 is partially correlated with CI expression, suggesting that it could be just linked to a CI gene.

The reliability of the length of wing radial vein r(2/3) as a character for the morphological discrimination of the two potential arbovirus vectors Culexpipiens s.s. and Cx. torrentium from Germany was reassessed, after this character had been neglected for more than 40 years. Additionally, multivariate morphometric analyses were applied to evaluate wing shape variation between both species. Although high-throughput molecular tools are now available to differentiate the two species, a simple, low-cost routine alternative may be useful in the absence of a molecular laboratory, such as under semi-field conditions. A thin-plate splines transformation confirmed that primarily the shrinkage of vein r(2/3) is responsible for the wing differences between the two species. In the bivariate analysis, the r(2/3)/r3 indices of Cx. pipiens s.s. and Cx. torrentium were 0.185 and 0.289, respectively, resulting in a correct classification of more than 91% of all tested specimens. Using the absolute length of vein r(2/3) alone still allowed for more than 90% accurate discrimination. Furthermore, classification accuracy of linear discriminant analysis exceeded 97%.

We evaluate three trapping methods for their effectiveness at capturing Culexpipiens and Culex torrentium, both enzootic vectors of bird-associated viruses in Europe. The comparisons, performed in two regions in Sweden, were among CDC traps baited with carbon dioxide, gravid traps, and ovitraps baited with hay infusion. The proportions of the two Culex species in a catch differed between trap types, with CDC traps catching a lower proportion of Cx. torrentium than both gravid traps and ovitraps. Between gravid traps and ovitraps, there was no difference in the proportions of the two species. The results indicate that Cx. torrentium may go undetected or underestimated compared to Cx. pipiens when using carbon dioxide baited CDC traps. The new insight of trap bias presented here adds an important dimension to consider when investigating these vectors of bird-associated viruses in the field.

The extensive use of insecticides to control vector populations has lead to the widespread development of different mechanisms of insecticide resistance. Mutations that confer insecticide resistance are often associated to fitness costs that prevent them from spreading to fixation. In vectors, such fitness costs include reductions in preimaginal survival, adult size, longevity, and fecundity. The most commonly invoked explanation for the nature of such pleiotropic effects of insecticide resistance is the existence of resource-based trade-offs. According to this hypothesis, insecticide resistance would deplete the energetic stores of vectors, reducing the energy available for other biological functions and generating trade-offs between insecticide resistance and key life history traits. Here we test this hypothesis by quantifying the energetic resources (lipids, glycogen, and glucose) of larvae and adult females of the mosquito Culexpipiens L. resistant to insecticides through two different mechanisms: esterase overproduction and acetylcholinesterase modification. We find that, as expected from trade-off theory, insecticide resistant mosquitoes through the overproduction of esterases contain on average 30% less energetic reserves than their susceptible counterparts. Acetylcholinesterase-modified mosquitoes, however, also showed a significant reduction in energetic resources (20% less). We suggest that, in acetylcholinesterase-modified mosquitoes, resource depletion may not be the result of resource-based trade-offs but a consequence of the hyperactivation of the nervous system. We argue that these results not only provide a mechanistic explanation for the negative pleiotropic effects of insecticide resistance on mosquito life history traits but also can have a direct effect on the development of parasites that depend on the vector's energetic reserves to fulfil their own metabolic needs.

Background. Outbreaks of the West Nile virus infection were reported in Morocco in 1996, 2003, and 2010. Culexpipiens was strongly suspected as the vector responsible for transmission. In the North center of Morocco, this species has developed resistance to synthetic insecticides. There is an urgent need to find alternatives to the insecticides as natural biocides. Objective. In this work, the insecticidal activity of the extract of the local plant Nerium oleander, which has never been tested before in the North center of Morocco, was studied on larval stages 3 and 4 of Culexpipiens. Methods. Biological tests were realized according to a methodology inspired from standard World Health Organization protocol. The mortality values were determined after 24 h of exposure and LC50 and LC90 values were calculated. Results. The extract had toxic effects on the larvae of culicid mosquitoes. The ethanolic extract of Nerium oleander applied against the larvae of Culexpipiens has given the lethal concentrations LC50 and LC90 in the order of 57.57 mg/mL and 166.35 mg/mL, respectively. Conclusion. This investigation indicates that N. oleander could serve as a potential larvicidal, effective natural biocide against mosquito larvae, particularly Culexpipiens. PMID:26640701

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) is now endemic in California across a variety of ecological regions that support a wide diversity of potential avian and mammalian host species. Because different avian hosts have varying competence for WNV, determining the blood-feeding patterns of Culex (Diptera: Culicidae) vectors is a key component in understanding the maintenance and amplification of the virus as well as tangential transmission to humans and horses. We investigated the blood-feeding patterns of Culex tarsalis Coquillett and members of the Culexpipiens L. complex from southern to northern California. Nearly 100 different host species were identified from 1,487 bloodmeals, by using the mitochondrial gene cytochrome c oxidase I (COI). Cx. tarsalis fed on a higher diversity of hosts and more frequently on nonhuman mammals than did the Cx. pipiens complex. Several WNV-competent host species, including house finch and house sparrow, were common bloodmeal sources for both vector species across several biomes and could account for WNV maintenance and amplification in these areas. Highly competent American crow, western scrub-jay and yellow-billed magpie also were fed upon often when available and are likely important as amplifying hosts for WNV in some areas. Neither species fed frequently on humans (Cx. pipiens complex [0.4%], Cx. tarsalis [0.2%]), but with high abundance, both species could serve as both enzootic and bridge vectors for WNV. PMID:22897051

Reproductive incompatibilities called cytoplasmic incompatibilities are known to affect a large number of arthropod species and are mediated by Wolbachia, a maternally transmitted microorganism. The crossing relationships between strains of potential hosts define their incompatibility types and it is generally assumed that differences between strains of Wolbachia induce different crossing types. Among all the described host species, the mosquito, Culexpipiens, displays the greatest variability of cytoplasmic incompatibility crossing types. We analysed mitochondrial and bacterial DNA variability in Culexpipiens in order to investigate some possible causes of incompatibility crossing type variability. We sequenced fragments of the ftsZ gene, and the A + T-rich control region of the mtDNA. We also sequenced the second subunit of the mitochondrial cytochrome oxidase (COII) gene, in Culexpipiens and a closely related species, C. torrentium, in order to verify the usefulness of the A + T-rich region for the present purposes. No variability was found in the Wolbachia ftsZ gene fragment, and very limited variation of the mitochondrial marker whatever the compatibility type or the origin of the host. A low variability was found in the A + T-rich region and comparison of divergence of the A + T-rich region and COII gene between C. pipiens and C. torrentium did not reveal any special constraints affecting this region. In contrast to observations in other host species, variability of incompatibility crossing types is not due to multiple infections by distantly related Wolbachia strains. PMID:9061971

Background: The aim of this study was to design pictorial key and taxonomic literature of Culexpipiens complex in Iran. Methods: Larvae were collected using standard dipping methods in 13 randomly selected areas of Bushehr, Hamedan, Kerman, Khorasan-e-Razavi, Khuzistan, Mazandaran, Tehran, Sistan and Baluchistan and Yazd Provinces from April 2009 to October 2010. The data were analyzed using SPSS Ver. 11.5. Results: Culexpipiens larvae were identified based on the Seta 1 of the abdominal segments III–IV in north and central parts of Iran. This diagnostic character had some variation among the Cx. quinquefasciatus collected from south of the country. The identification value of intersection of costa, subcosta and bifurcation of R2+3 of female veins, was calculated as 90–100 % for Cx. pipiens. This diagnostic character was varied among the Cx. quinquefasciatus specimens. The male genitalia found as the main characters to distinguish of Cx. quinquefasciatus from Cx. pipiens. Conclusion: It is necessary more studies on the behavior and genetic variations of Cx. pipiens complex in Iran. PMID:27308288

Larvae of the Culexpipiens complex (Cx. pipiens and Cx. torrentium) were abundant in two highly polluted pools receiving sewage sludge in Uppsala, Sweden (early August through late September 1985). The water was characterized by high BOD, and high ion concentration of Cu, Fe, Al and much suspended matter. Maximum larval number at the pool surface area was 26.1/ml. The ratio between species was studied and Cx. torrentium comprised ca. 20% at the peak of abundance. Some egg rafts showed no embryogeny.

Field studies were conducted in southwestern Virginia to determine the ovipositional preferences of Culex restuans and Culexpipiens by using ovitraps and gravid traps baited with selected infusions. For the ovitrap collections, 4 different infusions (manure, hay, grass, and rabbit chow) were used. Significant differences among infusions were detected on most sample dates for both species. For 3 of the first 4 wk of collections, the manure infusion collected significantly more Cx. restuans than all the other infusions. The hay and grass infusions collected the majority of the egg rafts during weeks 5-9. Cx. pipiens egg rafts were absent from the first 3 wk of collections. Of the remaining 6 wk, 4 showed significant differences in attractiveness of infusions, with the hay and grass infusions preferred by Cx. pipiens. Two infusions, manure and hay, were used for the gravid trap experiment and both Cx. restuans and Cx. pipiens data were combined for analysis. Only the first 2 wk showed significance, with manure being preferred over hay in both weeks. In later collections, the relative attractiveness of the hay infusion increased. A seasonal shift in infusion preference may be related to incubation temperature during preparation of the infusions. New infusions were prepared each week and incubation was done outside. Increased attractiveness of the hay infusion coincided with higher average temperatures in July and August. Hay infusion was very effective for trapping both Cx. pipiens and Cx. restuans in southwestern Virginia and is more convenient to use than manure. However, cool outside temperatures in the early season may interfere with the fermentation process and thus incubation should be done for a longer time or brought indoors.

The insecticidal activities of essential oil extracts from leaves, flowers and roots of aromatic plants against fourth-instar larvae of the mosquito Culexpipiens molestus Forskal were determined. Extracts of Foeniculum vulgare Mill were the most toxic, followed by those of Ferula hermonis Boiss, Citrus sinensis Osbeck, Pinus pinea L, Laurus nobilis L and Eucalyptus spp with LC50 values of 24.5, 44.0, 60.0, 75.0, 117.0 and 120.0 mg litre(-1), respectively. Combination tests between the LC50 and the maximum sub-lethal concentration (MSLC) were determined. Over 20 major components were identified in extracts from each plant species tested. Five essential oils and nine pure components were studied for their repellency against mosquito bites. Terpineol and 1,8-cineole were the most effective against Culexpipiens molestus bites offering complete protection for 1.6 and 2 h, respectively.

The southern house mosquito, Culexpipiens quinquefasciatus (Diptera: Culicidae), was introduced accidentally to Hawaii in 1826 (van Riper et al. 1986). There it eventually became the vector of avian malaria, Plasmodium relictum, a disease that severely limits the size and distribution of endemic forest bird populations in Hawaii (Atkinson et al. 1995). Cx.p. quinquefasciatus has a circumtropical distribution and is also the vector for human diseases such as lymphatic filariasis and several encephalitis.

Background Schmallenberg virus (SBV), an arboviral pathogen of ruminants, emerged in northern Europe during 2011 and has subsequently spread across a vast geographic area. While Culicoides biting midges (Diptera: Ceratopogonidae) have been identified as a biological transmission agent of SBV, the role of mosquitoes (Diptera: Culicidae) as potential vectors has not been defined beyond small-scale field collections in affected areas. Culexpipiens L. are one of the most widespread mosquitoes in northern Europe; they are present on farms across the region and have previously been implicated as vectors of several other arboviruses. We assessed the ability of three colony lines of Cx. pipiens, originating from geographically diverse field populations, to become fully infected by SBV using semi-quantitative real-time RT-PCR (sqPCR). Findings Two colony lines of Cx. pipiens were created in the UK (‘Brookwood’ and ‘Caldbeck’) from field collections of larvae and pupae and characterised using genetic markers. A third strain of Cx. pipiens from CVI Wageningen, The Netherlands, was also screened during experiments. Intrathoracic inoculation of the Brookwood line resulted in infections after 14 days that were characterised by high levels of RNA throughout individuals, but which demonstrated indirect evidence of salivary gland barriers. Feeding of 322 individuals across the three colony lines on a membrane based infection system resulted in no evidence of full dissemination of SBV, although infections did occur in a small proportion of Cx. pipiens from each line. Conclusions/Significance This study established two novel lines of Cx. pipiens mosquitoes of UK origin in the laboratory and subsequently tested their competence for SBV. Schmallenberg virus replication and dissemination was restricted, demonstrating that Cx. pipiens is unlikely to be an epidemiologically important vector of the virus in northern Europe. PMID:26291533

Females of Culexpipiens autogenicus generally have two refractory periods: post-emergence refractory period (during the first 48 hrs. after emergence) and post-insemination refractory period. Males of Culexpipiens autogenicus can inseminate 2 to 3 females. After the 4th insemination he can no longer completely fulfill his reproductory role. Therefore the female can inseminated by another male.

Background Japanese encephalitis virus (JEV) is the causative agent of Japanese encephalitis, the leading cause of viral encephalitis in Asia. JEV transmission cycle involves mosquitoes and vertebrate hosts. The detection of JEV RNA in a pool of Culexpipiens caught in 2010 in Italy raised the concern of a putative emergence of the virus in Europe. We aimed to study the vector competence of European mosquito populations, such as Cx. pipiens and Aedes albopictus for JEV genotypes 3 and 5. Findings After oral feeding on an infectious blood meal, mosquitoes were dissected at various times post-virus exposure. We found that the peak for JEV infection and transmission was between 11 and 13 days post-virus exposure. We observed a faster dissemination of both JEV genotypes in Ae. albopictus mosquitoes, when compared with Cx. pipiens mosquitoes. We also dissected salivary glands and collected saliva from infected mosquitoes and showed that Ae. albopictus mosquitoes transmitted JEV earlier than Cx. pipiens. The virus collected from Ae. albopictus and Cx. pipiens saliva was competent at causing pathogenesis in a mouse model for JEV infection. Using this model, we found that mosquito saliva or salivary glands did not enhance the severity of the disease. Conclusions In this study, we demonstrated that European populations of Ae. albopictus and Cx. pipiens were efficient vectors for JEV transmission. Susceptible vertebrate species that develop high viremia are an obligatory part of the JEV transmission cycle. This study highlights the need to investigate the susceptibility of potential JEV reservoir hosts in Europe, notably amongst swine populations and local water birds. PMID:28085881

Culexpipiens is the most cosmopolitan mosquito of the Pipiens Assemblage. By studying the nature of interactions between this species and microorganisms common to its breeding environment we can unravel important pitfalls encountered during development. We tested the survival rate of larval stages, pupae and adults of a Cx. pipiens colony exposed to a variety of microorganisms in laboratory conditions and assessed the transmission to offspring (F1) by those organisms that secured development up to adulthood. Three complementary experiments were designed to: 1) explore the nutritional value of yeasts and other microorganisms during Cx. pipiens development; 2) elucidate the transstadial transmission of yeast to the host offspring; and 3) to examine the relevance of all these microorganisms in female choice for oviposition-substratum. The yeast Saccharomyces cerevisiae proved to be the most nutritional diet, but despite showing the highest survival rates, vertical transmission to F1 was never confirmed. In addition, during the oviposition trials, none of the gravid females was attracted to the yeast substratum. Notably, the two native bacterial strains, Klebsiella sp. and Aeromonas sp., were the preferred oviposition media, the same two bacteria that managed to feed neonates until molting into 2nd instar larvae. Our results not only suggest that Klebsiella sp. or Aeromonas sp. serve as attractants for oviposition habitat selection, but also nurture the most fragile instar, L1, to assure molting into a more resilient stage, L2, while yeast proves to be the most supportive diet for completing development. These experiments unearthed survival traits that might be considered in the future development of strategies of Cx. pipiens control. These studies can be extended to other members of the Pipiens Assemblage. PMID:27055276

Background: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases. Methods: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culexpipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene. Results: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world. Conclusion: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure. PMID:27308293

Background: During recent years transmission of Dirofilaria immitis (dog heart worm) by Culexpipiens and West Nile virus have been reported from Iran. The present study was preformed for evaluating the susceptibility status of Cx. pipiens collected from capital city of Tehran, Iran. Methods: Four Insecticides including: DDT 4%, Lambdacyhalothrin 0.05%, Deltamethrin 0.05% and Cyfluthrin 0.15 % according to WHO standard methods were used for evaluating the susceptibility status of Cx. pipiens from Tehran moreover For comparison susceptibility status a Laboratory strain also was used. Bioassay data were analyzed using Probit program. The lethal time for 50% and 90% mortality (LT50 and LT90) values were calculated from regression line. Results: The susceptibility status of lab strain of Cx. pipiens revealed that it is susceptible to Lambdacyhalothrin, Deltamethrin, Cyfluthrin and resistant to DDT. Moreover cyfluthrin with LT50=36 seconds and DDT with LT50=3005 seconds had the least and most LT50s. Field population was resistance to all tested insecticides and DDT yielded no mortality. Conclusion: Highly resistance level against all WHO recommended imagicides were detected in field populations. We suggest more biochemical and molecular investigations to detect resistance mechanisms in the field population for further decision of vector control. PMID:28032100

The primary mosquito species associated with underground stormwater systems in the United States are the Culexpipiens complex species. This group represents important vectors of West Nile virus (WNV) throughout regions of the continental U.S. In this study, we designed a mathematical model and compared it with surveillance data for the Cx. pipiens complex collected in Beaufort County, South Carolina. Based on the best fit of the model to the data, we estimated parameters associated with the effectiveness of public health insecticide (adulticide) treatments (primarily pyrethrin products) as well as the birth, maturation, and death rates of immature and adult Cx. pipiens complex mosquitoes. We used these estimates for modeling the spread of WNV to obtain more reliable disease outbreak predictions and performed numerical simulations to test various mosquito abatement strategies. We demonstrated that insecticide treatments produced significant reductions in the Cx. pipiens complex populations. However, abatement efforts were effective for approximately one day and the vector mosquitoes rebounded until the next treatment. These results suggest that frequent insecticide applications are necessary to control these mosquitoes. We derived the basic reproductive number (ℜ0) to predict the conditions under which disease outbreaks are likely to occur and to evaluate mosquito abatement strategies. We concluded that enhancing the mosquito death rate results in lower values of ℜ0, and if ℜ0<1, then an epidemic will not occur. Our modeling results provide insights about control strategies of the vector populations and, consequently, a potential decrease in the risk of a WNV outbreak.

We investigated the role of heavy rain on container-inhabiting mosquito (Diptera: Culicidae) populations, and how different species may have adapted to such conditions. Rains were created with a rain simulator calibrated to natural rain intensities in the habitats of two important vector species: Aedes aegypti (L.) from northern Thailand and Culexpipiens L. from New York state, USA. Immature stages of Ae. aegypti were able to resist the flushing effect of rain better than Cx. pipiens. This difference was most dramatic during the pupal stage. Fourth instars of Ae. aegypti were not affected by flushing when exposed for longer rain intervals (30 versus 60 min) or at a colder water temperature (24 versus 16 degrees C). In contrast, significantly more Cx. pipiens larvae flushed out with longer rain exposure. Warmer water temperatures also increased the proportion of Cx. pipiens flushed out, but mostly at the longest exposure time. Container position (tilted at a 7 degrees angle or level) did not affect proportions of fourth instars flushed out for both species. More accurate models of vector-borne diseases can be developed by incorporating the described effects of rain on container-breeding mosquito populations. Such models may provide more realistic assessments of disease risk and ensure optimal use of limited financial resources of mosquito control agencies.

The status of genes conferring resistance to organophosphate and carbamate insecticides has been examined in Culexpipienspipiens mosquitoes sampled in Algeria. Presence of overproduced esterases was sporadic, but acetylcholinesterase-1 resistant alleles were observed in almost all samples. We focused our study on the AChE1 G119S substitution characterized in almost all samples, mostly at the heterozygous state. A genetic test revealed the presence of ace-1 duplication associating a susceptible and a resistant ace-1 copy. Molecular characterization showed a high occurrence of ace-1 duplication with six distinct duplicated alleles out of four samples. The inferred frequency of duplicated allele suggests that it is replacing the single resistant G119S allele. Finally, we discuss the mechanism at the origin of these duplicated haplotypes and their consequences on the management of insecticide resistance.

Background: Malaria is still a health problem in Iran. There are several vector control activities, including Indoor Residual spraying, using insecticide treated nets and larviciding including Temephos. In addition nuisance mosquitos are prevalent in the urban areas. So that evaluation of this species to larvicide will provide a clue for management of vector control activities. Methods: Two mosquito species were used in this study: Anopheles stephensi were collected from Kazeroun and Culexpipiens from Tehran, capital of Iran. All the tests were carried out according to the WHO method. All the test kis was provided by WHO. Results: Results showed a LC50= 0.0523 and LC90=0.3822 mg/l for An. stephensi. The figure for Cx. pipiens was 0.1838 and 0.8505 mg/l respectively. Conclusion: monitoring of insecticide resistance to Temephos should be evaluated regularly for management of vector control. PMID:28032103

The vector competence and bionomics of Culexpipiens form pipiens L. and Cx. pipiens f. molestus Forskäl were evaluated for populations from the Sacramento Valley. Both f. pipiens and f. molestus females became infected, produced disseminated infections, and were able to transmit West Nile virus. Form molestus females also transmitted West Nile virus vertically to egg rafts and F1 progeny, whereas f. pipiens females only transmitted to egg rafts. Culexpipiens complex from urban Sacramento blood-fed on seven different avian species and two mammalian species. Structure analysis of blood-fed mosquitoes identified K = 4 genetic clusters: f. molestus, f. pipiens, a group of genetically similar hybrids (Cluster X), and admixed individuals. When females were exposed as larvae to midwinter conditions in bioenvironmental chambers, 85% (N = 79) of aboveground Cx. pipiens complex females and 100% (N = 34) of underground f. molestus females did not enter reproductive diapause.

In East Asia, 4 forms of the Culexpipiens complex have been confirmed. A form pipiens s. s. (anautogeous pipiens) has been confirmed only in westernmost China. A temperate form pallens and a subtropical and tropical form quinquefasciatus are connected with intermediates in morphology and many aspects of ecology, but their difference is rather clear in climatic adaptation traits. The distribution of a form molestus overlaps with pallens and, in Taiwan, quinquefasciatus, and, in East Asia, this form requires artificial underground habitats for its persistence. The origin and interrelation of 3 forms other than pipiens s. s. are considered from ecological aspects, especially climatic adaptation. A hypothesis is presented that molestus was originally a form having adapted to the Mediterranean climate in the western Palaearctic, secondarily colonized artificial underground habitats, and reached East Asia in the early 20th century by ships from North America. At present, it is difficult to assign pallens with certainty to either the old or new groups of the Manchurian mosquito fauna. Three hypotheses about the interrelation between pallens and quinquefasciatus are compared, and the strengths and problems of each hypothesis are indicated. Finally, a map to show the distribution of the forms of the Culexpipiens complex before it was extensively changed by humans is presented as an initial trial. The Culexpipiens problems now troubling humans are largely human-made problems.

In Morocco, Culexpipiens plays a role in the high annoyance experienced by most urban cities, suburban and rural areas, especially since it was strongly suspected as the most likely vector in the transmission of West Nile virus epidemics that have hit Morocco in 1996. Chemical insecticides are generally the way in which they use the programs against harmful mosquitoes and disease vectors. However, the repeated and excessive use of these products regularly led to the emergence of the phenomenon of insect resistance. At the center of Morocco, information on the susceptibility or resistance to insecticides in mosquitoes (larvae and adults) vectors of diseases or pests, are almost nonexistent. This article reports the results of studies conducted between 2007 and 2010 with sensitivity tests WHO on larvae local populations of Culexpipiens collected in three lodging in the city of Fez, towards the insecticide mostly used by hygienic services: temephos. Five concentrations of insecticide (0.0025 mg/l, 0.005 mg/l, 0.0125 mg/l, 0.025 mg/l, 0.0625 mg/l) in addition to control, were used to determine the LC50 and LC 90 of Culexpipiens species towards temephos. Sensitivity tests were carried out at the entomology unit and monitoring of insect sensitivity towards insecticides installed at the Regional Diagnostic Laboratory Epidemiological and Environmental Hygiene (LRDEHM), Fez, under the Regional Directorate of Health in Fes Boulemane Region. The LC50 and LC90, concentrations corresponding to 50 and 90% mortality were determined graphically, by the linear relationship between the decimal logarithm of insecticide concentrations (x-axis) and the percentage of mortality transformed into probit values (ordinate) on logarithmic gausso paper. Resistance rates were determined on the basis of the sensitivity of a reference strain (S-Lab). The bioassay results affirmed the presence of resistance in larvae Culexpipiens towards temephos and that this species has also equally

Fresh leaves of Myrtus communis collected from different localities in Greece, were subjected to hydrodistillation and the oils obtained were analyzed by GC-FID and GC-MS. The analyses showed mainly quantitative differences, with the monoterpenes myrtenyl acetate, α-pinene, 1,8-cineole, and linalool, along with limonene, dominating the majority of the analyzed Myrtle oils. The evaluation of the larvicidal activity of the samples against Culexpipiens biotype molestus mosquito showed that all tested samples exhibited moderate to weak toxicity, with cultivated M. communis subsp. communis oil being the most active.

The Culexpipiens complex includes two widespread mosquito vector species, Cx. pipiens and Cx. quinquefasciatus. The distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. However, their distribution range overlaps in certain areas and interspecific hybridization has been documented. Genetic introgression between these species may have epidemiological repercussions for West Nile virus (WNV) transmission. Bayesian clustering analysis based on multilocus genotypes of 12 microsatellites was used to determine levels of hybridization between these two species in Macaronesian islands, the only contact zone described in West Africa. The distribution of the two species reflects both the islands' biogeography and historical aspects of human colonization. Madeira Island displayed a homogenous population of Cx. pipiens, whereas Cape Verde showed a more intriguing scenario with extensive hybridization. In the islands of Brava and Santiago, only Cx. quinquefasciatus was found, while in Fogo and Maio high hybrid rates (∼40%) between the two species were detected. Within the admixed populations, second-generation hybrids (∼50%) were identified suggesting a lack of isolation mechanisms. The observed levels of hybridization may locally potentiate the transmission to humans of zoonotic arboviruses such as WNV. PMID:22957190

In this study of the mosquito, Culexpipiens, we examined the impact of multiple bouts of dehydration and rehydration on survival, depletion of metabolic reserves and egg production in both non-diapausing and diapausing females. Mosquitoes provided with access to sugar during rehydration survived longer than those allowed to rehydrate without sugar, and their survival was similar to that of mosquitoes of the same age that were not dehydrated. Among mosquitoes not provided with sugar, each dehydration bout reduced the mosquito's dry mass – an effect likely to be due to the utilization of carbohydrates and lipid reserves. The toll on glycogen and lipid reserves is likely to be especially costly for diapausing mosquitoes that are dependent on these stored reserves for winter survival. Egg production in both non-diapausing and post-diapausing C. pipiens was also reduced in response to multiple bouts of dehydration. Although egg quality was not compromised, the number of eggs produced was reduced. Both non-diapausing and diapausing females can compensate for the nutrient loss due to dehydration by sugar feeding but the opportunity to feed on sugar is likely to be rarely available in the overwintering habitat of diapausing females, thus the impact of dehydration may be especially pronounced in overwintering populations of C. pipiens. PMID:20675546

Wolbachia are maternally inherited endosymbionts that can invade arthropod populations through manipulation of their reproduction. In mosquitoes, Wolbachia induce embryonic death, known as cytoplasmic incompatibility (CI), whenever infected males mate with females either uninfected or infected with an incompatible strain. Although genetic determinants of CI are unknown, a functional model involving the so-called mod and resc factors has been proposed. Natural populations of Culexpipiens mosquito display a complex CI relationship pattern associated with the highest Wolbachia (wPip) genetic polymorphism reported so far. We show here that C. pipiens populations from La Réunion, a geographically isolated island in the southwest of the Indian Ocean, are infected with genetically closely related wPip strains. Crossing experiments reveal that these Wolbachia are all mutually compatible. However, crosses with genetically more distant wPip strains indicate that Wolbachia strains from La Réunion belong to at least five distinct incompatibility groups (or crossing types). These incompatibility properties which are strictly independent from the nuclear background, formally establish that in C. pipiens, CI is controlled by several Wolbachia mod/resc factors.

Despite the presence of Culex (Cx.) pipiens mosquitoes and circulation of West Nile virus (WNV), WNV outbreaks have so far not occurred in northern Europe. The species Cx. pipiens consists of two morphologically identical biotypes, pipiens and molestus, which can form hybrids. Until now, population dynamic studies of Cx. pipiens have not differentiated between biotypes and hybrids at the European scale, nor have they used comparative surveillance approaches. We therefore aimed to elucidate the relative abundance of Cx. pipiens biotypes and hybrids in three habitat types at different latitudes across Europe, using two different surveillance traps. BG-Sentinel and Mosquito-Magnet Liberty Plus traps were placed in three habitat types (farms, peri-urban, wetlands), in three European countries (Sweden, The Netherlands, Italy). Collected Cx. pipiens mosquitoes were identified to biotype with real-time PCR. Both trap types collected equal ratios of the biotypes and their hybrids. From northern to southern latitudes there was a significant decrease of pipiens and an increase of molestus. Habitat types influenced the relative ratios of biotypes and hybrids, but results were not consistent across latitudes. Our results emphasize the need to differentiate Cx. pipiens to the biotype level, especially for proper future WNV risk assessments for Europe. PMID:27870890

In this study we probe the molecular events underpinning diapause observed in overwintering females of Culexpipiens. Using suppressive subtractive hybridization (SSH) we have identified 40 genes that are either upregulated or downregulated during this seasonal period of dormancy. Northern blot hybr...

Mosquito populations are sensitive to long-term variations in climate and short-term variations in weather. Mosquito abundance is a key determinant of outbreaks of mosquito-borne diseases, such as West Nile virus (WNV). In this work, the short-term impact of weather conditions (temperature and precipitation) on Culexpipiens L.-Culex restuans Theobald mosquito abundance in Peel Region, Ontario, Canada, was investigated using the 2002-2009 mosquito data collected from the WNV surveillance program managed by Ontario Ministry of Health and Long-Term Care and a gamma-generalized linear model. There was a clear association between weather conditions (temperature and precipitation) and mosquito abundance, which allowed the definition of threshold criteria for temperature and precipitation conditions for mosquito population growth. A predictive statistical model for mosquito population based on weather conditions was calibrated using real weather and mosquito surveillance data, and validated using a subset of surveillance data. Results showed that WNV vector abundance on any one day could be predicted with reasonable accuracy from relationships with mean degree-days >9 degrees C over the 11 preceding days, and precipitation 35 d previously. This finding provides optimism for the development of weather-generated forecasting for WNV risk that could be used in decision support systems for interventions such as mosquito control.

The widespread and improper use of pyrethroid insecticides, such as deltamethrin, has resulted in the evolution of resistance in many mosquito species, including Culexpipiens pallens. With the development of high-throughput sequencing, it is possible to massively screen pyrethroid resistance-associated gene. In this study, we used Illumina-Solexa transcriptome sequencing to identify genes that are expressed differently in deltamethrin-susceptible and -resistant strains of Culexpipiens pallens as a critical knowledge base for further studies. A total of 4,961,197,620 base pairs and 55,124,418 reads were sequenced, mapped to the Culex quinquefasciatus genome and assembled into 17,679 known genes. We recorded 1,826 significantly differentially expressed genes (DEGs). Among them, 1,078 genes were up-regulated and 748 genes were down-regulated in the deltamethrin-resistant strain compared to -susceptible strain. These DEGs contained cytochrome P450s, cuticle proteins, UDP-glucuronosyltransferases, lipases, serine proteases, heat shock proteins, esterases and others. Among the 1,826 DEGs, we found that the transcriptional levels of CYP6AA9 in the laboratory populations was elevated as the levels of deltamethrin resistance increased. Moreover, the expression levels of the CYP6AA9 were significantly higher in the resistant strains than the susceptible strains in three different field populations. We further confirmed the association between the CYP6AA9 gene and deltamethrin resistance in mosquitoes by RNA interfering (RNAi). Altogether, we explored massive potential pyrethroid resistance-associated genes and demonstrated that CYP6AA9 participated in the pyrethroid resistance in mosquitoes. PMID:26377942

The primary mosquito species associated with underground stormwater systems in the United States are the Culexpipiens complex species. This group represents important vectors of West Nile virus (WNV) throughout regions of the continental U.S. In this study, we designed a mathematical model and compared it with surveillance data for the Cx. pipiens complex collected in Beaufort County, South Carolina. Based on the best fit of the model to the data, we estimated parameters associated with the effectiveness of public health insecticide (adulticide) treatments (primarily pyrethrin products) as well as the birth, maturation, and death rates of immature and adult Cx. pipiens complex mosquitoes. We used these estimates for modeling the spread of WNV to obtain more reliable disease outbreak predictions and performed numerical simulations to test various mosquito abatement strategies. We demonstrated that insecticide treatments produced significant reductions in the Cx. pipiens complex populations. However, abatement efforts were effective for approximately one day and the vector mosquitoes rebounded until the next treatment. These results suggest that frequent insecticide applications are necessary to control these mosquitoes. We derived the basic reproductive number (ℜ0) to predict the conditions under which disease outbreaks are likely to occur and to evaluate mosquito abatement strategies. We concluded that enhancing the mosquito death rate results in lower values of ℜ0, and if ℜ0<1, then an epidemic will not occur. Our modeling results provide insights about control strategies of the vector populations and, consequently, a potential decrease in the risk of a WNV outbreak. PMID:25268229

Both ornithophilic mosquito species, Culexpipiens s.l. (L.) and Culex torrentium (Martini, 1925), occur sympatric in temperate Europe. They are presumed to be primary vectors of West Nile and Sindbis viruses. Differentiation of these morphologically similar Culex species is essential for evaluation of different vector roles, for mosquito surveillance and integrated control strategies. Cx. torrentium has been neglected or erroneously determined as Cx. pipiens s.l. in some previous studies, because only males of both species can be diagnosed reliably by morphology. Thus, knowledge about species abundance, geographical distribution, breeding site preferences and the zoonotic risk assessment is incomplete also in Poland. In Wrocław area (Silesian Lowland), besides typical urban breeding sites, huge sewage irrigation fields provide suitable breeding conditions for Culex species. They are also inhabited by 180 resident and migratory bird species serving as potential virus reservoirs. In this study, morphology of larvae and males as well as species diagnostic enzyme markers, namely adenylate kinase (AK) and 2-hydroxybutyrate dehydrogenase (HBDH), were used to discriminate Cx. pipiens s.l. and Cx. torrentium. In a total of 650 Culex larvae from 24 natural and artificial breeding sites, Cx. pipiens s.l. had a proportion of 94.0% and Cx. torrentium only 6.0%. It could be shown that both species are well adapted to various breeding site types like ditches, catch basins, flower pots and buckets with diverse water quality. Cx. torrentium preferred more artificial water containers in urban surrounding (12% species proportion), whereas in semi-natural breeding sites, Cx. torrentium was rare (3%). In 12 of 24 breeding sites, larvae of both species have been found associated.

Since April 1963, the susceptibility to insecticides of larvae of Culexpipiens fatigans has been extensively studied by the WHO Filariasis Research Unit in Rangoon, Burma, as part of a programme of research aimed at establishing which compound or group of compounds could be used to control the vector of filariasis. Larvae from various localities in Rangoon showed wide variations in susceptibility to chlorinated hydrocarbons. There were also seasonal variations and it was difficult to standardize test conditions. Nevertheless, resistance to this group of compounds, especially DDT, sufficient to prevent control has been clearly demonstrated. As C. p. fatigans has never been subjected to intensive control by insecticides in Rangoon, the results confirm the findings made elsewhere that this mosquito is naturally resistant to chlorinated hydrocarbons. The larvae are, however, susceptible to organophosphorus compounds, especially fenthion and parathion. PMID:4230023

Mosquitoes in the Culexpipiens complex, Cx. p. pipiens L. (Cpp) and Cx. p. quinquefasciatus Say (Cpq), are morphologically similar and important vectors of the West Nile and St. Louis Encephalitis viruses in the US. They hybridize when found in sympatry, which could facilitate the transfer of adva...

Background Knowledge on the temporal dynamics of host/vector/parasite interactions is a pre-requisite to further address relevant questions in the fields of epidemiology and evolutionary ecology of infectious diseases. In studies of avian malaria, the natural history of Plasmodium parasites with their natural mosquito vectors, however, is mostly unknown. Methods Using artificial water containers placed in the field, we monitored the relative abundance of parous females of Culexpipiens mosquitoes during two years (2010–2011), in a population in western Switzerland. Additionally, we used molecular tools to examine changes in avian malaria prevalence and Plasmodium lineage composition in female C. pipiens caught throughout one field season (April-August) in 2011. Results C. pipiens relative abundance varied both between years and months, and was associated with temperature fluctuations. Total Plasmodium prevalence was high and increased from spring to summer months (13.1-20.3%). The Plasmodium community was composed of seven different lineages including P. relictum (SGS1, GRW11 and PADOM02 lineages), P. vaughani (lineage SYAT05) and other Plasmodium spp. (AFTRU5, PADOM1, COLL1). The most prevalent lineages, P. vaughani (lineage SYAT05) and P. relictum (lineage SGS1), were consistently found between years, although they had antagonistic dominance patterns during the season survey. Conclusions Our results suggest that the time window of analysis is critical in evaluating changes in the community of avian malaria lineages infecting mosquitoes. The potential determinants of the observed changes as well as their implications for future prospects on avian malaria are discussed. PMID:24499594

Mosquitoes such as those in the Culexpipiens complex are important vectors of disease. This study was conducted to genetically characterize Cx. pipiens complex populations in the state of Colorado, USA, and to determine the number of genetic clusters represented by the data. Thirteen populations located among four major river basins were sampled (n = 597 individuals) using a panel of 14 microsatellites. The lowest-elevation sites had the highest Expected Heterozygosity (H(E)) values (range 0.54-0.65). AMOVA results indicated the presence of statistically significant amounts of variation within each level when populations were analyzed as one group or when they were grouped either by river basin or by their position on the east or west side of the Rocky Mountains. Most pairwise F(ST) values were significant via permutation test (range 0-0.10), with the highest values from comparisons with Lamar, in southeast CO. A neighbor joining tree based on Cavalli-Sforza and Edwards's chord distances was consistent with the geographic locations of populations, as well as with the AMOVA results. There was a significant isolation by distance effect, and the cluster analysis resolved five groups. Individuals were also assayed with an additional microsatellite marker, Cxpq78, proposed to be monomorphic in Cx. pipiens but polymorphic in the closely related but biologically distinct species Cx. quinquefasciatus. Low frequencies (≤3%) of Cx. quinquefasciatus alleles for this marker were noted, and mostly confined to populations along the Interstate 25 corridor. Pueblo was distinct in that it had 10% Cx. quinquefasciatus alleles, mostly of one allele size. The degree of population genetic structure observed in this study is in contrast with that of Cx. tarsalis, the other major vector of WNV in the western U.S., and likely reflects the two species' different dispersal strategies.

This study evaluated the toxicity of Colocasia esculenta leaves extract on 3rd, 4th instars larvae and pupae of Culexpipiens. Bioassays showed that the 3rd instar larvae was the most susceptible to the different concentrations of extract, where the LC50 after 48 hr. post-exposure was 79.41, 109.65 & 141.25 for the 3rd, 4th instars larvae and pupal stage respectively. The histo-pathological effects of C. esculenta leaves extract on midgut regions and gastric caeca of the 3rd instar larvae were studied. When larvae were treated with 100 ppm of C. esculenta extract, all larvae developed dramatic pathological lesions especially Malpighian tubules were extensively affected. The midgut cells showed morphological deviation from normal ones, through slightly apical degenerated (lysis) of epithelial cells. The epithelial cells with extensive cellular microvilli were shrinkage, the nuclei showed pyknotic characteristic and the peritrophic membrane was appeared discontinuation in compared to control. When the 3rd larval instar was exposed to extract 400 ppm, the epithelial cells, adipose fabric and muscles were extensively affected. Also, the gastric caeca was affected obviously. These observation and alterations in cells of Cx. pipiens larvae are related to the dangerous effect of C. esculent leaves extract.

A species of Amblyospora-infecting neurones of Culexpipiens is described. Diplokaryotic meronts, which divided by binary fission, were distinguished at the electron microscope level by their unthickened plasma membranes. Sporonts with an electron-dense surface coat gave rise to eight uninucleate sporoblasts within a sporophorous vesicle, cytoplasmic division occurring at the quadrinucleate or octonucleate stages. Indications that nuclear fusion and chromosome reorganization occurred in merogony and sporogony were obtained by light microscopy but meiosis was not detected at the ultrastructural level. Spores were typical of Amblyospora, being ovoid when fresh, truncate when stained, and having an exospore of two membranous layers subtended by a thick amorphous layer, an electron-lucent endospore, an anisofilar polar filament, and a polaroplast comprised of an anterior region of close-packed lamellae and a posterior region of expanded sacs. The metabolic products in the sporophorous vesicle took the form of large globules, small globules with electron-dense borders, and fine granules. These were depleted in mature sporophorous vesicles, though a surface layer of fine granules on the spores may have been derived from them. Many stages were degenerate and it is suggested that C. pipiens may be an accidental host in which the parasite could develop suboptimally in nervous tissue only. Infections in larvae hatched from eggs in the laboratory indicate that vertical transmission occurs.

The couch potato (cpo) cDNA that we cloned from the northern house mosquito, Culexpipiens, encodes the C-terminus containing a highly conserved RNA recognition motif (RRM). Protein structure prediction indicates a canonical RRM structure with a βαββαβ topological structure. Northern blots indicate a single mRNA band over 9.49 kb, and Southern blot analysis suggests that the cpo gene contains large introns. Highest expression was noted in first instar larvae and pupae. A comparison of nondiapausing (long daylength) and diapausing (short daylength) adult females showed no difference immediately at adult eclosion, but by day 7 and thereafter, expression of cpo was much higher in diapausing adults. When 2-month old diapausing females were transferred from short daylength to diapausing-terminating conditions of long daylength and high temperature, expression of cpo declined. Similarly, when a topical application of JH III was used to terminate diapause abundance of the cpo transcript declined. Consistent with observations in Drosophila melanogaster and several other species levels of cpo in C. pipiens are influenced by the diapause program, although the direction of change is not the same in all species.

Previous studies in our laboratory found that the extract from seeds of Amorpha fruticosa in the Leguminosae family had lethal effects against mosquito larvae, and an insecticidal compound amorphigenin was isolated. In this study, the inhibitory effects of amorphigenin against the mitochondrial complex I of Culexpipiens pallens (Diptera: Culicidae) were investigated and compared with that of rotenone. The results showed that amorphigenin and rotenone can decrease the mitochondrial complex I activity both in vivo and in vitro as the in vivo IC50 values (the inhibitor concentrations leading to 50% of the enzyme activity lost) were determined to be 2.4329 and 2.5232 μmol/L, respectively, while the in vitro IC50 values were 2.8592 and 3.1375 μmol/L, respectively. Both amorphigenin and rotenone were shown to be reversible and mixed-I type inhibitors of the mitochondrial complex I of Cx. pipiens pallens, indicating that amorphigenin and rotenone inhibited the enzyme activity not only by binding with the free enzyme but also with the enzyme-substrate complex, and the values of KI and KIS for amorphigenin were determined to be 20.58 and 87.55 μM, respectively, while the values for rotenone were 14.04 and 69.23 μM, respectively. PMID:26307964

The continuous culture of mosquitoes is a costly endeavor for vector biology laboratories. In addition to the resources that must be committed to colony maintenance, biological costs, including genetic drift and accidental colony loss, also can occur. Although alternatives do exist, their application to mosquitoes is limited. Mosquito cryopreservation remains elusive, and many important species lack a well-defined diapause. Previously, we demonstrated that cold storing nondiapausing mated adult females of the northern house mosquito, Culexpipiens L. resulted in a nearly four-fold increase in longevity when measured at the LT50, allowing for cold storage for up to 10 wk. In the current study, we used sugar feeding during cold storage to significantly improve cold storage longevity. At 6 degrees C, the LT50 of cold stored females was 23 wk, and 100% mortality was not realized until 43 wk. Cold-stored females did exhibit reduced fecundity, but egg production returned to normal levels within two generations. These results suggest that cold storage without diapause induction is a viable option for Cx. pipiens, and with the addition of sugar feeding, a colony could be maintained with less than two generations per year.

Nowadays a number of endemic mosquito species are known to possess vector abilities for various diseases, as e.g. the sibling species Culexpipiens and Culex torrentium. Due to their morphological similarity, ecology, distribution and vector abilities, knowledge about these species' population structure is essential. Culicidae from 25 different sampling sites were collected from March till October 2012. All analyses were performed with aligned cox1 sequences with a total length of 658 bp. Population structure as well as distribution patterns of both species were analysed using molecular methods and different statistical tests like distance based redundancy analysis (dbDRA), analysis of molecular variances (AMOVA) or McDonald & Kreitman test and Tajima's D. Within both species, we could show a genetic variability among the cox1 fragment. The construction of haplotype networks revealed one dominating haplotype for Cx. pipiens, widely distributed within Germany and a more homogeneous pattern for Cx. torrentium. The low genetic differences within Cx. pipiens could be a result of an infection with Wolbachia which can induce a sweep through populations by passively taking the also maternally inherited mtDNA through the population, thereby reducing the mitochondrial diversity as an outcome of reproductive incompatibility. Pairwise population genetic differentiation (FST) ranged significantly from moderate to very great between populations of Cx. pipiens and Cx. torrentium. Analyses of molecular variances revealed for both species that the main genetic variability exists within the populations (Cx. pipiens [88.38%]; Cx. torrentium [66.54%]). Based on a distance based redundancy analysis geographical origin explained a small but significant part of the species' genetic variation. Overall, the results confirm that Cx. pipiens and Cx. torrentium underlie different factors regarding their mitochondrial differentiation, which could be a result of endosymbiosis, dispersal

Dirofilaria immitis is one of the most frequently detected mosquito-transmitted zoonotic filarioid nematode in mammals in Europe, being canine dirofilariosis a major animal health problem, endemic in the Mediterranean area. This study, focused on Southwest Spain, in order to bring new insights into (i) the epidemiology of Dirofilaria spp., (ii) the species of Culicid vectors possibly involved in their transmission and (iii) the genetic variability of those potential vectors. A total of 881 adult female mosquitoes from 11 different species, were captured during 2012-2013, and detection of filarioid DNA was attempted by PCR using specific primers (ITS-2 and COI), followed by DNA sequencing. In a single Culexpipiens specimen D. immitis DNA was detected both in the head-thorax and abdomen sections. Filarioid nematode DNA was also detected in eight additional Cx. pipiens specimens also in both the thorax and the abdomen, but analysis of sequence data did not allow unambiguous assignment of any of the obtained sequences to a previously defined species. All Cx. pipiens with filarioid DNA were individually analysed by CQ11 to discriminate between pipiens, molestus, and hybrid forms. Besides, rDNA ITS-2 sequence analysis revealed the presence of haplotype H1 and H2 of Cx. pipiens. To our knowledge this study revealed, for the first time in Spain, the occurrence of likely mature infection of D. immitis in Cx. pipiens, as well as with other yet uncharacterized nematodes, supporting its role as a potential vector of these filarids.

The attract-and-kill strategy is a new pest management technique that presupposes the intelligent combination of an attracting agent (e.g. pheromone) and a killing agent (e.g. insecticide). In the present study, the potential combination of the microencapsulated synthetic oviposition pheromone 6-acetoxy-5-hexadecanolide with an insecticide has been tested. Initially, polyurea microcapsules containing 6-acetoxy-5-hexadecanolide, the synthetic mixture of diastereomers of the oviposition pheromone of the mosquito species Culex quinquefasciatus Say (Diptera: Culicidae), were studied. Laboratory bioassays were performed to confirm the bioactivity of the microencapsulated pheromone on the oviposition activity of Culexpipiens L. biotype molestus Førskal (Diptera: Culicidae) with the aim of determining the optimum dose for oviposition response. Its effect was dose dependent, revealing an optimum dose of 300 mg of dried microcapsules. Attractancy over time was also studied. The microencapsulated pheromone was found to be sufficiently attractive to gravid female mosquitoes for a period of 40 days. Finally, the combination of the synthetic pheromone with the control agent temephos showed both an acceptable oviposition activity and sufficient larvicidal effect.

Complex interactions between microbial residents of mosquitoes and arboviruses are likely to influence many aspects of vectorial capacity and could potentially have profound effects on patterns of arbovirus transmission. Such interactions have not been well studied for West Nile virus (WNV; Flaviviridae, Flavivirus) and Culex spp. mosquitoes. We utilized next-generation sequencing of 16S ribosomal RNA bacterial genes derived from Culexpipiens Linnaeus following WNV exposure and/or infection and compared bacterial populations and broad immune responses to unexposed mosquitoes. Our results demonstrate that WNV infection increases the diversity of bacterial populations and is associated with up-regulation of classical invertebrate immune pathways including RNA interference (RNAi), Toll, and Jak-STAT (Janus kinase-Signal Transducer and Activator of Transcription). In addition, WNV exposure alone, without the establishment of infection, results in similar alterations to microbial and immune signatures, although to a lesser extent. Multiple bacterial genera were found in greater abundance in WNV-exposed and/or infected mosquitoes, yet the most consistent and notable was the genus Serratia. PMID:26516902

The introduction of Aedes albopictus (Skuse) in the United States has been associated with declines in abundance of resident mosquito species, presumably because of resource competition, as larvae of Ae. albopictus have been illustrated as superior competitors under certain resource conditions. We evaluated the hypothesis that varying the type and ratio of two food resources (Foxtail grass: American elm) alters the competitive outcome of Ae. albopictus and Culexpipiens (L.). We measured survivorship, development time, size, and adult longevity, and estimated the population growth index (A') of populations raised both alone and in equal number with the interspecific competitor, across five ratios of the two food resources. Competition was asymmetric with Ae. albopictus, the superior competitor across all resource treatments; however, the competitive advantage Ae. albopictus had over Cx. pipiens was reduced as grass became the predominant resource. With elm as the predominant resource, the population growth index (A') for both Ae. albopictus and Cx. pipiens was lower in intraspecific and interspecific competition treatments, respectively. The treatments also impacted adult life history, as life spans of both Ae. albopictus and Cx. pipiens varied when they emerged from larval conditions with different resource and competition treatments. We discuss the possible differences in the two species' efficiencies in exploiting the two resource types. Despite some resource conditions alleviating the competitive effects of Ae. albopictus on Cx. pipiens, competition remained asymmetric; thus, additional mechanisms are likely operating under field conditions when the two species coexist.

The aim of our study was to assess the potential of Macrocyclops albidus as a biological control agent against the 1st and 2nd instars of Culexpipiens and Aedes albopictus. Under laboratory conditions of prey saturation (50 1st instars/copepod), an average of 58.98% of Cx. pipiens and 54.99% of Ae. albopictus larvae were killed by 1 copepod in 24 h. Trials run in big drums containing 200 liters of water showed that the M. albidus population, inoculated in April, efficiently controlled the mosquito population for the entire season. The predator was particularly effective against Ae. albopictus, as only 2 larvae of this species were found in the treated drums, compared to 814 larvae in untreated control drums throughout the study period. No difference was observed in the control efficacy between the 2 initial densities of copepods used. The reduction in Ae. albopictus density in the drums with 100 and 500 M. albidus with respect to the control drums was 99.90 ± 0.35% and 100.0 ± 0.0%, respectively. For Cx. pipiens, the reduction in density was 88.69 ± 13.51% and 84.65% in drums inoculated with 100 and 500 copepods, respectively. Macrocyclops albidus populations survived through the winter and continued to keep the mosquito population under control during the 2008 season. The M. albidus population developed very well both in drums placed in sunny and shaded areas and proved to be tolerant to both high (summer) and low (winter) temperatures. Trials performed on M. albidus survival in catch basins showed that after a few weeks, the copepod population dramatically decreased and subsequently disappeared. The main problem for copepod survival in catch basins seemed to be the low oxygen tension and accumulation of toxic substances, rather than copepods being flushed out in heavy rainfall episodes. During the period when copepods were present, they maintained the mosquito population under control; their partial disappearance from the catch basins, however, would

Background Culexpipiens/restuans mosquitoes are important vectors for a variety of arthropod borne viral infections. In this study, the associations between 20 years of mosquito capture data and the time lagged environmental quantities daytime length, temperature, precipitation, relative humidity and wind speed were used to generate a predictive model for the population dynamics of this vector species. Methods Mosquito population in the study area was represented by averaged time series of mosquitos counts captured at 6 sites in Cook County (Illinois, USA). Cross-correlation maps (CCMs) were compiled to investigate the association between mosquito abundances and environmental quantities. The results obtained from the CCMs were incorporated into a Poisson regression to generate a predictive model. To optimize the predictive model the time lags obtained from the CCMs were adjusted using a genetic algorithm. Results CCMs for weekly data showed a highly positive correlation of mosquito abundances with daytime length 4 to 5 weeks prior to capture (quantified by a Spearman rank order correlation of rS = 0.898) and with temperature during 2 weeks prior to capture (rS = 0.870). Maximal negative correlations were found for wind speed averaged over 3 week prior to capture (rS = −0.621). Cx. pipiens/restuans population dynamics was predicted by integrating the CCM results in Poisson regression models. They were used to simulate the average seasonal cycle of the mosquito abundance. Verification with observations resulted in a correlation of rS = 0.899 for daily and rS = 0.917 for weekly data. Applying the optimized models to the entire 20-years time series also resulted in a suitable fit with rS = 0.876 for daily and rS = 0.899 for weekly data. Conclusions The study demonstrates the application of interval lagged weather data to predict mosquito abundances with a feasible accuracy, especially when related to weekly Cx. pipiens

The aim of this study was to model the impact of temperature on the timing of the seasonal shift in relative proportion of Culex restuans Theobald and Culexpipiens L. in Illinois. The temporal pattern of West Nile virus (WNV) and St. Louis encephalitis virus transmission in the midwest exhibits a late summer to early fall peak in activity, which parallels the temporal increase in the abundance of Cx. pipiens. The daily number of egg rafts oviposited by each species has been monitored at multiple surveillance sites in Urbana-Champaign in central Illinois for more than 13 years. The time when the two Culex species are in equal abundance (crossover) varies considerably from year to year. Our investigation of several thermal measures indicated that this variation was related in large part to climatic conditions with warmer (cooler) temperatures correlated to earlier (later) crossover dates. Models based on degree days and the number of days in which the daily maximum temperature exceeded an upper temperature threshold explained more than 60% of the variance in crossover dates. In contrast, models based on the number of days in which the daily minimum temperature exceeded a lower temperature threshold explained no more than 52% of the variance. An evaluation of these models demonstrated that they provide relatively simple and accurate estimates of crossover date from daily temperature data, a necessary component for developing an overall climatic index for the risk of WNV transmission in Illinois.

Culexpipiens is the mosquito that vectors West Nile Virus and other human-pathogenic flavivruses in North America. In response to shortened day length and lower temperatures, female Cx. pipiense prepares for the diapause by actively feeding on carbohydrates to increase the biosynthesis of glycogen and lipid to store energy for overwintering. The effect of feeding different carbohydrates on glycogen and lipid biosynthesis in diapausing mosquitoes was investigated in vivo using 13C solid-state NMR. Diapause-destined adult females and nondiapausing counterparts after adult eclosion were fed with three different carbohydrate sources for 7 days: 1) 10% sucrose, 2) 10% D-[13C6]glucose, and 3) 1% D-[13C6]glucose co-provisioned with 10% sucrose. NMR measurements show that sucrose and glucose are metabolized differently in diapausing mosquitoes. Mosquitoes fed on sucrose primarily accumulate glycogen with increased branching structures, but less of lipids. In contrast, mosquitoes fed exclusively on glucose show accumulation of both glycogen and lipid with increased aliphatic chain length. Glucose is exclusively metabolized for the biosynthesis of triacylglyceride when mosquitoes were co-fed with sucrose. Our findings provide novel insights into the insect carbohydrate metabolism that governs glycogen and lipid biosynthesis during diapause, which is fundamental for the insect survival during inimical environments.

Chlorfenapyr is a promising pyrrole insecticide with a unique mechanism of action that does not confer cross-resistance to neurotoxic insecticides. The effect of chlorfenapyr on pyrethorid-resistant Culexpipiens pallens Coq (Diptera: Culicidae) has not been fully investigated under laboratory conditions. In this study, cypermethrin-resistant C. p. pallens exhibited 376.79-fold and 395.40-fold increase in resistance to cypermethrin compared with susceptible strains after exposure for 24 and 48h, respectively. Larvae and adults were tested for susceptibility using dipping, topical, and impregnated paper methods as recommended by the WHO. No cross-resistance to chlorfenapyr was found. Increased mortality was apparent between 48 and 72h, indicating a slow rate of toxic activity. Synergism experiments with piperonyl butoxide (PBO) showed an antagonistic effect on chlorfenapyr toxicity. Mixtures of chlorfenapyr and cypermethrin could therefore provide additional benefits over either insecticide used alone. Mixtures of 5ng/ml chlorfenapyr and 500ng/ml cypermethrin exhibited a slight synergistic effect on cypermethrin-resistant mosquitoes (3.33, 6.84 and 2.34% after 24, 48 and 72h exposure, respectively. This activity was lost when the chlorfenapyr concentration was increased to 10 or 20ng/ml. Chlorfenapyr showed quite good results for pyrethroid-resistant C. p. pallens, and could improve public health by reducing the occurrence of mosquito bites and subsequently protecting against transmission of lymphatic filariasis and Japanese encephalitis.

Ethanolic extracts from the Ginkgo biloba L. exocarp from the Chinese ginkgo were assayed against larvae of three strains of Culexpipiens pallens Coquillett. The chemical compositions were detected using a Hewlett-Packard 6890/5973 mass spectrometric detector. The larvicidal bioassay was carried out according to the recommendations of the World Health Organization. The analysis of the essential oil of ginkgo exocarp showed that its major components are ginkgo acid (85.3%) and ginkgo phenolic (5.69%). The larvicidal bioassay showed that extracts of ginkgo exocarp have LC50 of 18.6, 12.7, and 25.0 mg/liter for deltamethrin-susceptible, deltamethrin-resistant, and field strains, respectively. The acute toxicity concentrations of the ginkgo extracts that killed 50% (LD50) of Wistar rats within 2 wk and young carp within 96 h were 4947.2 mg/kg and 557.9 mg/liter, respectively. These results are promising in creating new, effective, and affordable approaches to mosquito control.

Culexpipiens is the mosquito that vectors West Nile Virus and other human-pathogenic flavivruses in North America. In response to shortened day length and lower temperatures, female Cx. pipiense prepares for the diapause by actively feeding on carbohydrates to increase the biosynthesis of glycogen and lipid to store energy for overwintering. The effect of feeding different carbohydrates on glycogen and lipid biosynthesis in diapausing mosquitoes was investigated in vivo using 13C solid-state NMR. Diapause-destined adult females and nondiapausing counterparts after adult eclosion were fed with three different carbohydrate sources for 7 days: 1) 10% sucrose, 2) 10% D-[13C6]glucose, and 3) 1% D-[13C6]glucose co-provisioned with 10% sucrose. NMR measurements show that sucrose and glucose are metabolized differently in diapausing mosquitoes. Mosquitoes fed on sucrose primarily accumulate glycogen with increased branching structures, but less of lipids. In contrast, mosquitoes fed exclusively on glucose show accumulation of both glycogen and lipid with increased aliphatic chain length. Glucose is exclusively metabolized for the biosynthesis of triacylglyceride when mosquitoes were co-fed with sucrose. Our findings provide novel insights into the insect carbohydrate metabolism that governs glycogen and lipid biosynthesis during diapause, which is fundamental for the insect survival during inimical environments. PMID:27853281

Alterations in gene expression in the midgut of female Culexpipiens quinquefasciatus exposed to blood meals containing 6.8 logs plaque-forming units/mL of West Nile virus (WNV) were studied by fluorescent differential display. Twenty-six different cDNAs exhibited reproducible differences after feeding on infected blood. Of these, 21 cDNAs showed an increase in expression, and 5 showed a decrease in expression as a result of WNV presence in the blood meal. GenBank database searches showed that one clone with increased expression, CQ G12A2, shares 94% identity with a leucine-rich repeat-containing protein from Cx. p. quinquefasciatus and 32% identity to Toll-like receptors from Aedes aegypti. We present the first cDNA clone isolated from female Cx. p. quinquefasciatus midgut tissue whose expression changes on exposure to WNV. This cDNA represents a mosquito gene that is an excellent candidate for interacting with WNV in Cx. p. quinquefasciatus and may play a role in disease transmission. PMID:19635880

Laboratory experiments to determine the duration of the immature stages of Culexpipiens fatigans were carried out because such information is important from the point of view of control by larvicides. At a temperature of 25.1°C±0.7°C the mean incubation period is 27.11±0.57 hours. Females spend a longer time in the pupal stage than males (34.16±0.74 hours and 32.95±0.75 hours, respectively, at 28.6°C±0.8°C; there is no 24-hour pupating or emerging rhythm. The duration of larval life is longer for the female (135.3±4.4 hours) than for the male (118.4±2.4 hours). Larvae that take a long time to pupate also take a long time to emerge. Withholding of food for a few hours from first-stage larvae increases the duration of larval life but does not affect that of pupal life. These observations on the differences between the sexes in the duration of larval and pupal life are in agreement with observations made on Aedes aegypti in Uganda. PMID:4227199

Two amino acid substitutions in acetylcholinesterase 1 (AChE1), G119S and F290V, are responsible for resistance to organophosphate and carbamate insecticides in Culexpipiens mosquitoes. These mutations generate very different levels of insensitivity to insecticide inhibitors. We described here a biochemical method that rapidly identifies AChE1 variants (susceptible, G119S and F290V, named S, R and V, respectively) present in individual mosquitoes. We investigated the frequency of AChE1 phenotypes in 41 field samples collected around the Mediterranean Sea. F290V substitution was found only in 15 samples and at low frequency, whereas G119S was highly spread in all samples. However, seven V distinct alleles were identified whereas only one R allele was present. The [V] enzymatic phenotype was never observed alone, and the V allele was always found associated with the susceptible and/or G119S AChE1 ([VS], [VR] or [VRS] phenotypes). Furthermore, we showed the presence of duplicated alleles, associating a susceptible and a V copy of the ace-1 gene, in most individuals analyzed for its presence. Evolutionary forces driving the large number of F290V ace-1 alleles and their low frequency in Mediterranean countries are discussed.

Resistance to insecticides was monitored on Culexpipiens quinquefasciatus mosquitoes collected in twelve localities of La Réunion, a geographically isolated island of the Indian Ocean. This mosquito is of medical concern in the region as a known vector for filariasis and a potential vector for West Nile and Rift Valley Fever viruses. Our bioassays indicated the presence of resistance to all tested insecticides, i.e. organochlorides, organophosphates and pyrethroids. A molecular investigation revealed a higher frequency of resistance genes in the coastal areas compared to elevated rural sites, probably reflecting the different nature of insecticide pressures together with the genetic cost of resistance alleles. A simple molecular test was developed to detect Rdl allele, encoding a gamma-aminobutyric acid (GABA) receptor resistant to dieldrin. Unexpectedly high Rdl frequencies were recorded over the whole island, despite this insecticide having been banned for over 15 years. This resistant allele was also detected for the first time in two samples of Aedes albopictus, a species recently involved in severe Chikungunya epidemics on the island. Rdl selection in these two mosquito species discloses current insecticide pressures in urban areas, from unknown origins, that should be taken into account to develop vector control strategies.

The mechanism of DDT resistance in Culexpipiens fatigans is poorly understood. Earlier studies indicated that the dehydrochlorination of DDT does not explain resistance in this species. Studies on the role of lipids as a mechanism of resistance included the estimation of lipid content and the determination of the proportions of different classes of lipids in the larvae of susceptible and resistant strains. There was no evidence of any correlation between the lipid content and DDT resistance in this species and the proportions of neutral lipids, phospholipids and fatty acids of different strains did not indicate any consistent correlation with DDT resistance. Within one strain, the larvae containing the higher amount of lipids were able to resist better the toxic effect of DDT. Analysis of fatty acids of the larvae that survived and died as a result of treatment with DDT did not reveal any difference. Neither p,p′-DDT nor o,p′-DDT at sublethal concentration affected the lipids of the larvae of susceptible and resistant strains. PMID:5310957

The effect of non-selecting doses of DDT on the induction of resistance was investigated in the house mosquito, Culexpipiens. Sublethal doses of DDT were applied to the adults in each generation, at levels which just fell short of causing mortality in these adults. A susceptible strain, a DDT-resistant strain, and a slightly DDT-tolerant strain were so treated for 6-7 generations.It was found that the treatment initially caused approximately 25% of the ovaries to degenerate, and reduced the proportion of females that fed and oviposited. This reduction in biotic potential became aggravated in successive generations of the DDT-resistant and DDT-tolerant strains, which failed to show any material increase in resistance level. In the susceptible strain, however, the biotic potential became enhanced, and considerable resistance was developed. It was concluded that the increase of resistance in this strain was due to hidden selection, of eggs in the ovary, and of females which failed to oviposit.

Most mosquito species display host preferences that are a crucial determinant of the transmission rate of mosquito-borne pathogens. Although a transgenic approach, based on driving genes for zoophily into vector populations, has been advocated as a malaria control strategy by the World Health Organization since 1982, the genes involved in mosquito host choice remain poorly understood. Culexpipiens pallens Coquillet mosquitoes were artificially selected for two different host preferences in a specially designed experimental enclosure. Of 3,035 mosquitoes obtained from larvae and pupae collected from the wild (the F0 generation), 27% preferentially fed on pigeons and 16% fed on mice. Following artificial selection for these host preferences over successive generations, the percentage of mosquitoes that preferred to feed on pigeons or mice gradually increased, eventually stabilizing at ∼55 and 34%, respectively, after the sixth generation. Intergenerational differences in host preferences were significant (P

Culex (Culex) pipiens s.l. (Diptera: Culicidae) comprises two distinct biotypes, pipiens ('rural') and molestus ('urban'), both of which are thought to have differing capacities due to different host preferences. To better understand West Nile encephalitis epidemiology and improve risk assessment, local distinction between these forms is essential. This study assesses phenotypic variation at larval and adult stages of 'urban' and 'rural' biotypes of the species by complementary use of meristic, univariate and multivariate traits analyzed by traditional and geometric morphometrics. Third- and fourth-instar larvae from a broad area of the city of Novi Sad (Serbia) were collected and reared in the laboratory. After adult eclosion, the sex of each larva was recorded based on the sex of the corresponding adult. Examination of the association between variations of larval traits revealed contrasting variations regarding pecten spines vs. siphonal size and siphonal shape in the 'rural' biotype. Siphons of larvae collected in marshes and forest ecosystems outside urban areas were found to be the largest, but possessed the smallest number of pecten spines. In addition, statistically significant female-biased sexual dimorphism was observed in siphonal size, wing size and wing shape. Finally, we propose that an integrative approach is essential in delimitation of Cx. pipiens s.l. biotypes, since their differentiation was not possible based solely on larval and adult traits. Our findings shed light on the phenotypic plasticity important for population persistence in the changing environment of these medically important taxa.

Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI) resulting in abortive embryonic development. Among all the described host species, mosquitoes of the Culexpipiens complex display the highest variability of CI crossing types. Paradoxically, searches for polymorphism in Wolbachia infecting strains and field populations hitherto failed or produced very few markers. Here, we show that an abundant source of the long-sought polymorphism lies in WO prophage sequences present in multiple copies dispersed in the genome of Wolbachia infecting C. pipiens (wPip). We identified up to 66 different Wolbachia variants in C. pipiens strains and field populations and no occurrence of superinfection was observed. At least 49 different Wolbachia occurred in Southern Europe C. pipiens populations, and up to 10 different Wolbachia were even detected in a single population. This is in sharp contrast with North African and Cretan samples, which exhibited only six variants. The WO polymorphism appeared stable over time, and was exclusively transferred maternally. Interestingly, we found that the CI pattern previously described correlates with the variability of Gp15, a prophage protein similar to a bacterial virulence protein. WO prophage sequences thus represent variable markers that now open routes for approaching the molecular basis of CI, the host effects, the structure and dynamics of Wolbachia populations. PMID:16615218

Polymerase chain reaction (PCR) is often used to detect microorganisms, pathogens, or both, including the reproductive parasite Wolbachia pipientis (Rickettsiales: Anaplasmataceae), in mosquitoes. Natural populations of Culexpipiens L. (Diptera: Culicidae) mosquitoes are infected with one or more strains of W. pipientis, and crosses between mosquitoes harboring different Wolbachia strains provide one of the best-known examples of cytoplasmic incompatibililty (CI). When we used PCR to monitor Wolbachia in the Buckeye strain of Culexpipiens, and in a Wolbachia-cured sister colony obtained by tetracycline treatment, we noted false negative PCR reactions with DNA samples from infected mosquitoes; these results were inconsistent with direct microscopic observation of Wolbachia-like particles in gonads dissected from mosquitoes in the same population. Assays with diluted template often improved detection of positive samples, suggesting that DNA prepared from whole mosquitoes contained an inhibitor of the PCR reaction. We reconciled discrepancies between PCR and microscopy by systematic measurement of the PCR reaction in the presence of an internal standard. Mosquito decapitation before DNA extraction restored the reliability of the PCR reaction, allowing accurate determination of Wolbachia infection status in infected and tetracycline-cured mosquito populations, consistent with microscopic examination. Using PCR primers based on the Tr1 gene, we confirmed that the Wolbachia infection in the Buckeye strain of Culexpipiens belongs to the genotype designated wPip1. Finally, to explore more widely the distribution of PCR inhibitors, we demonstrated that DNA isolated from the cricket, Acheta domesticus (L.); the beetle, Tenebrio molitor L.; the honey bee, Apis mellifera L.; and the mosquito, Anopheles punctipennis Say also contained PCR inhibitors. These results underscore the importance of measuring the presence of inhibitors in PCR templates by using a known positive

Diapause is a critical eco-physiological adaptation for winter survival in the West Nile Virus vector, Culexpipiens, but little is known about the molecular mechanisms that distinguish diapause from non-diapause in this important mosquito species. We used Illumina RNA-seq to simultaneously identify and quantify relative transcript levels in diapausing and non-diapausing adult females. Among 65,623,095 read pairs, we identified 41 genes with significantly different transcript abundances between these two groups. Transcriptome divergences between these two phenotypes include genes related to juvenile hormone synthesis, anaerobic metabolism, innate immunity and cold tolerance. PMID:27128578

MicroRNAs (miRNAs) regulate gene expression and biological processes including embryonic development, innate immunity and infection in many species. Emerging evidence indicates that miRNAs are involved in drug resistance. However, little is known about the relationship between the miRNAs and insecticide resistance in mosquitos. Here, we reported that conserved miR-278-3p and its target gene are critical for pyrethroid resistance in Culexpipiens pallens. We found that CYP6AG11 is the target of miR-278-3p, through bioinformatic analysis and experimental verification. The expression level of miR-278-3p was lower, whereas the level of CYP6AG11 was higher in deltamethrin-resistant strain, which were detected using qRT-PCR. We also found that CYP6AG11 was regulated by miR-278-3p via a specific target site with the 3′UTR by luciferase reporter assay. In addition, overexpression of CYP6AG11 in the mosquito C6/36 cells showed better prolification than the cells with empty vector when treated by deltamethrin at different concentrations. Moreover, the overexpression of miR-278-3p through microinjection led to a significant reduction in the survival rate, and the level of CYP6AG11 was simultaneously reduced. These results indicated that miR-278-3p could regulate the pyrethroid resistance through CYP6AG11. PMID:25420996

We investigated the genetic determinism of high chlorpyrifos resistance (HCR), a phenotype first described in 1999 in Culexpipiens mosquitoes surviving chlorpyrifos doses ⩾1 mg l−1 and more recently found in field samples from Tunisia, Israel or Indian Ocean islands. Through chlorpyrifos selection, we selected several HCR strains that displayed over 10 000-fold resistance. All strains were homozygous for resistant alleles at two main loci: the ace-1 gene, with the resistant ace-1R allele expressing the insensitive G119S acetylcholinesterase, and a resistant allele of an unknown gene (named T) linked to the sex and ace-2 genes. We constructed a strain carrying only the T-resistant allele and studied its resistance characteristics. By crossing this strain with strains harboring different alleles at the ace-1 locus, we showed that the resistant ace-1R and the T alleles act in strong synergy, as they elicited a resistance 100 times higher than expected from a simple multiplicative effect. This effect was specific to chlorpyrifos and parathion and was not affected by synergists. We also examined how HCR was expressed in strains carrying other ace-1-resistant alleles, such as ace-1V or the duplicated ace-1D allele, currently spreading worldwide. We identified two major parameters that influenced the level of resistance: the number and the nature of the ace-1-resistant alleles and the number of T alleles. Our data fit a model that predicts that the T allele acts by decreasing chlorpyrifos concentration in the compartment targeted in insects. PMID:26463842

OBJECTIVES To assess vector competence (infection, dissemination and transmission) of Culexpipiens quinquefasciatus for Florida (FL) West Nile virus (WNV) isolates. METHODS West Nile virus isolates (WN-FL-03: NY99 genotype; WN-FL-05-558, WN-FL-05-2186, WN-FL-05-510: WN02 genotype) collected from different regions of FL were used for vector competence experiments in Cx. p. quinquefasciatus from Alachua County and Indian River County in FL. Mosquitoes from both colonies were fed blood containing 7.9 ± 0.2 log10 plaque-forming units WNV/ml ± SE and incubated at 28 °C for 14 days. Vector competence, including rates of infection, dissemination, and transmission, was compared between colonies for WN-FL-03 using chi-squared. Virus titres in bodies, legs and saliva were compared using ANOVA. Daily measurements of in vitro replication of WNV isolates were evaluated in Vero cells so that a standardised virus dose for each isolate could be delivered to mosquitoes. RESULTS Infection and dissemination rates were high (≥95%) and not affected by isolate or colony (infection, P = 0.679; dissemination, P = 0.799). Transmission rates were low (≤20%), detected in one colony and affected by isolate (P = 0.008). Body and leg titres differed between isolates (body titre, P = 0.031; leg titre, P = 0.044) and colonies (body titre, P = 0.001; leg titre, P = 0.013) while saliva titre did not differ between isolates (P = 0.462). CONCLUSIONS Variation in vector competence of mosquito populations may be attributed, in part, to exposures to WNV with genetic differences leading to different rates of replication in mosquitoes. Evaluation of vector competence for different WNV isolates may help us understand vector–virus interactions and, hence, the role of vectors in complex virus transmission cycles in nature. PMID:24898274

We investigated the genetic determinism of high chlorpyrifos resistance (HCR), a phenotype first described in 1999 in Culexpipiens mosquitoes surviving chlorpyrifos doses ⩾1 mg l(-1) and more recently found in field samples from Tunisia, Israel or Indian Ocean islands. Through chlorpyrifos selection, we selected several HCR strains that displayed over 10 000-fold resistance. All strains were homozygous for resistant alleles at two main loci: the ace-1 gene, with the resistant ace-1(R) allele expressing the insensitive G119S acetylcholinesterase, and a resistant allele of an unknown gene (named T) linked to the sex and ace-2 genes. We constructed a strain carrying only the T-resistant allele and studied its resistance characteristics. By crossing this strain with strains harboring different alleles at the ace-1 locus, we showed that the resistant ace-1(R) and the T alleles act in strong synergy, as they elicited a resistance 100 times higher than expected from a simple multiplicative effect. This effect was specific to chlorpyrifos and parathion and was not affected by synergists. We also examined how HCR was expressed in strains carrying other ace-1-resistant alleles, such as ace-1(V) or the duplicated ace-1(D) allele, currently spreading worldwide. We identified two major parameters that influenced the level of resistance: the number and the nature of the ace-1-resistant alleles and the number of T alleles. Our data fit a model that predicts that the T allele acts by decreasing chlorpyrifos concentration in the compartment targeted in insects.

Background: The extracts of different parts of plants were found very effective against various pests. The aim of this research was to determine the insecticidal activity of fruit methanol extracts obtained from Melia azedarach (Meliaceae), Phoenix theophrasti (Arecaceae), Styphnolobium japonicum (Fabaceae) and Pyracantha coccinea (Rosaceae) against the larvae of Culexpipiens (Diptera: Culicidae). Methods: The fruits of test plants were collected from the Campus of Akdeniz University, Antalya, Turkey in 2013. A series of concentrations of the extracts ranging from 62.5–1000 ppm were tested against second instar larvae. Results: Only the extracts of Me. azedarach and Ph. theoprasti showed significant larvicidal activity against Cx. pipiens and the LC50 values of these extracts were found to be 169.48 and 220.60 ppm, respectively. This is the first research investigating the insecticidal or larvicidal activity of Ph. theophrasti, St. japonicum and Py. coccinea extracts on mosquitoes. Conclusion: The methanol extract of fruits of Me. azedarach and Ph. theophrasti showed significantly higher larvicidal activity against Cx. pipiens. PMID:28032112

The insecticidal effect of six commercially available plant oils was tested against 4th larval instars of Culexpipiens. Larvae were originally collected from Meit El-Attar, Qalyubia Governorate, Egypt, and then reared in the laboratory until F1 generation. The LC50 values were 32.42, 47.17, 71.37, 83.36, 86.06, and 152.94 ppm for fenugreek (Trigonella foenum-grecum), earth almond (Cyperus esculentus), mustard (Brassica compestris), olibanum (Boswellia serrata), rocket (Eruca sativa), and parsley (Carum ptroselinum), respectively. The tested oils altered some biological aspects of C. pipiens, for instance, developmental periods, pupation rates, and adult emergences. The lowest concentrations of olibanum and fenugreek oils caused remarkable prolongation of larval and pupal durations. Data also showed that the increase of concentrations was directly proportional to reduction in pupation rates and adult emergences. Remarkable decrease in pupation rate was achieved by mustard oil at 1000 ppm. Adult emergence was suppressed by earth almond and fenugreek oils at 25 ppm. In addition, the tested plant oils exhibited various morphological abnormalities on larvae, pupae, and adult stages. Consequently, fenugreek was the most potent oil and the major cause of malformation of both larval and pupal stages. Potency of the applied plant oils provided an excellent potential for controlling C. pipiens.

Mosquitoes are one of the most globally important insect pests and vectors of human pathogens, and their populations may be facilitated or inhibited by anthropogenic environmental change. Invasive plant species are an important management concern and environmental modifier in many ecosystems; these plant invasions have the potential to exacerbate or diminish mosquito populations. The purpose of this study was to assess potential effects of a highly invasive plant, Lonicera maackii, on a common mosquito species Culexpipiens L., which is an important pathogen vector in the United States. Three microcosm assays were conducted to determine the responses of C. pipiens life history attributes of larval survivorship, growth, and pupation when subjected to leachate from two native plant leaves (Platanus occidentalis and Acer saccharum) and both the leaves and flowers of L. maackii. Only C. pipiens larvae exposed to L. maackii leachate pupated and emerged as adults. However, in all three assays there were statistically significant differences in survivorship and body size change among treatments, and in each assay the highest survivorship and maximum larval size was found in the L. maackii leachate treatments, suggesting positive effects on certain life history traits. This study is one of the first to demonstrate the potential facilitative effect of this invasive plant species on an insect vector and suggests that plant invasion could have positive feedbacks into mosquito population dynamics and, ultimately, human disease.

Adult mosquitoes rely on ingestion of sugar from plants to survive, swarm and mate. Culexpipiens pallens Coguillett is the primary vector of lymphatic filariasis and epidemic encephalitis. Little is known about the effect of feeding on different sugar sources on the survivorship and fecundity of Cx. pipiens pallens. In the present study, newly emerged mosquitoes were exposed to several flowering plant and seed pod species with different olfactory preferences, and the survival times of mosquitoes exposed to these sugar sources were determined. The proportions of mosquitoes that ingested sugar from host plants were investigated by cold anthrone tests. The numbers of eggs per egg raft laid by mosquitoes were compared when they were provided with different sugar sources and one blood meal. The results revealed that feeding on different kinds of sugar sources significantly affected female and male mosquitoes' survival times. Cold anthrone tests indicated that the proportions of sugar-positive mosquitoes from different nutritional regimes within 24h corresponded to the preference rankings of Cx. pipiens pallens to these sugar sources, and rapid declines in the proportions of surviving individuals might be attributed to their insufficient ingestion of sugar from nutritional regimes. Feeding on different sugar sources strongly affected the proportions of engorged mosquitoes, and females that had fed on their preferred sugar sources laid more eggs than mosquitoes provided with less preferred sugar sources. The results would provide insights in developing mosquito control strategies that target the sugar feeding behavior of mosquitoes.

Two actin genes cloned from Culexpipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized acti...

The host blood-feeding patterns of mosquito vectors affects the likelihood of human exposure to zoonotic pathogens, including West Nile Virus (family Flaviviridae, genus Flavivirus, WNV). In Portugal, data are unavailable regarding the blood-feeding habits of common mosquito species, including Culexpipiens L., considered the primary vector of WNV to humans. The sources of bloodmeals in 203 blood-fed mosquitoes of nine species collected from June 2007 to November 2010 in 34 Portuguese counties were analyzed by sequencing cytochrome-b partial fragments. Cx. pipiens was the most common species collected and successfully analyzed (n = 135/78). In addition, blood-fed females of the following species were analyzed: Ochlerotatus caspius Pallas (n = 20), Culex theileri Theobald (n = 16), Anopheles maculipennis s.l. Meigen (n = 10), Culiseta longiareolata Macquart (n = 7), Aedes aegypti L. (n = 6), Culex perexiguus Theobald (n = 3), Culiseta annulata Schrank (n = 3), and Ochlerotatus detritus Haliday (n = 3). The Cx. pipiens mosquitoes fed predominantly on birds (n = 55/78, 70.5%), with a high diversity of avian species used as hosts, although human blood was identified in 18 specimens (18/78, 23.1%). No significant differences were found between the host-feeding patterns of blood-fed Cx. pipiens collected in residential and nonresidential habitats. The occurrence of human derived blood meals and the presence of a mix avian-human bloodmeal accordingly suggest this species as a potential vector of WNV. Therefore, in Portugal, Cx. pipiens may play a role both in the avian-to-avian enzootic WNV cycle and in the avian-to-mammal transmission. In this context, the identity of Cx. pipiens (considering the forms molestus and pipiens) and the potential consequence on feeding behavior and WNV transmission are discussed.

An acetylcholinesterase (AChE) gene, Ace.x, showing 93% identity of deduced amino acid sequence to Anopheles stephensi Ace has been cloned from a Culexpipiens strain homozygous for insensitive AChE (iAChE) mediated insecticide resistance. DNA sequence of genomic DNA clones identified exons 2-5. RFLP of six clones indicated four possible alleles. Linkage analysis located Ace.x to chromosome I, less than 0.8 centimorgans from the sex locus, whereas the locus conferring resistance was 2.0 centimorgans from plum-eye on chromosome II. Ace.1 coding for AChE1, which is associated with resistance, is therefore autosomal. We propose that Ace.x is the recently postulated Ace.2 coding for the biochemically distinct AChE2, which is not associated with resistance.

We examined the effect of simulated overwintering temperatures on West Nile (WN) virus replication in Culexpipiens L. derived from mosquitoes collected during the autumn 1999 WN epizootic in New York. The WN virus was a strain isolated from a dead crow also collected during this outbreak. Virus was recovered from most mosquitoes held exclusively at 26 degres C. In contrast, none of the mosquitoes held exclusively at the lower temperatures had detectable infections. When mosquitoes were transferred to 26 degrees C after being held at 10 degrees C for 21-42 d, infection and dissemination rates increased with increased incubation at 26 degrees C. Future studies involving the attempted isolation of WN virus from overwintering mosquitoes may benefit from holding the mosquitoes at 26 degrees C before testing for infectious virus.

Before maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culexpipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and approximately 98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction. PMID:14704183

West Nile fever (WNF) and Rift Valley fever (RVF) are emerging diseases causing epidemics outside their natural range of distribution. West Nile virus (WNV) circulates widely and harmlessly in the old world among birds as amplifying hosts, and horses and humans as accidental dead-end hosts. Rift Valley fever virus (RVFV) re-emerges periodically in Africa causing massive outbreaks. In the Maghreb, eco-climatic and entomologic conditions are favourable for WNV and RVFV emergence. Both viruses are transmitted by mosquitoes belonging to the Culexpipiens complex. We evaluated the ability of different populations of Cx. pipiens from North Africa to transmit WNV and the avirulent RVFV Clone 13 strain. Mosquitoes collected in Algeria, Morocco, and Tunisia during the summer 2010 were experimentally infected with WNV and RVFV Clone 13 strain at titers of 107.8 and 108.5 plaque forming units/mL, respectively. Disseminated infection and transmission rates were estimated 14–21 days following the exposure to the infectious blood-meal. We show that 14 days after exposure to WNV, all mosquito st developed a high disseminated infection and were able to excrete infectious saliva. However, only 69.2% of mosquito strains developed a disseminated infection with RVFV Clone 13 strain, and among them, 77.8% were able to deliver virus through saliva. Thus, Cx. pipiens from the Maghreb are efficient experimental vectors to transmit WNV and to a lesser extent, RVFV Clone 13 strain. The epidemiologic importance of our findings should be considered in the light of other parameters related to mosquito ecology and biology. PMID:22693557

West Nile fever (WNF) and Rift Valley fever (RVF) are emerging diseases causing epidemics outside their natural range of distribution. West Nile virus (WNV) circulates widely and harmlessly in the old world among birds as amplifying hosts, and horses and humans as accidental dead-end hosts. Rift Valley fever virus (RVFV) re-emerges periodically in Africa causing massive outbreaks. In the Maghreb, eco-climatic and entomologic conditions are favourable for WNV and RVFV emergence. Both viruses are transmitted by mosquitoes belonging to the Culexpipiens complex. We evaluated the ability of different populations of Cx. pipiens from North Africa to transmit WNV and the avirulent RVFV Clone 13 strain. Mosquitoes collected in Algeria, Morocco, and Tunisia during the summer 2010 were experimentally infected with WNV and RVFV Clone 13 strain at titers of 10(7.8) and 10(8.5) plaque forming units/mL, respectively. Disseminated infection and transmission rates were estimated 14-21 days following the exposure to the infectious blood-meal. We show that 14 days after exposure to WNV, all mosquito st developed a high disseminated infection and were able to excrete infectious saliva. However, only 69.2% of mosquito strains developed a disseminated infection with RVFV Clone 13 strain, and among them, 77.8% were able to deliver virus through saliva. Thus, Cx. pipiens from the Maghreb are efficient experimental vectors to transmit WNV and to a lesser extent, RVFV Clone 13 strain. The epidemiologic importance of our findings should be considered in the light of other parameters related to mosquito ecology and biology.

Different mixtures of monoterpenes (ketone, alcohol, and alkene) were loaded on paper discs and wax and their knockdown activities were evaluated against Culexpipiens adults. Some individual monoterpenes were also evaluated by residual toxicity technique. Citronella oil as a reference was also loaded separately or in combination with monoterpenes on paper discs and wax. The ketone monoterpenes mixture (camphor, menthone, carvone, and fenchone) on paper discs was the most active (KT50 = 17.20 min) followed by ketone monoterpenes with citronella oil (KT50 = 20.79 min) and citronella oil alone (KT50 = 28.72 min). Wax formulations proved that the ketone and alcohol (geraniol, thymol, and menthol) monoterpenes gave the most activity as knockdown (KT50 = 31.79 and 43.39 min, resp.). Alcohol monoterpenes formulation recorded KT50 = 43.39 min. Residual activity of tested individual monoterpenes reported that the menthol was more toxic than camphor and camphene. Generally, this study suggests that the monoterpenes have the properties, which make them used as eco-friendly compounds in the control programs of Cx. pipiens adult. The use of paper discs is more applicable than wax in the adulticidal formulations. PMID:27891154

Avian malaria studies have taken a prominent place in different aspects of evolutionary ecology. Despite a recent interest in the role of vectors within the complex interaction system of the malaria parasite, they have largely been ignored in most epidemiological studies. Epidemiology of the disease is however strongly related to the vector's ecology and behaviour, and there is a need for basic investigations to obtain a better picture of the natural associations between Plasmodium lineages, vector species and bird hosts. The aim of the present study was to identify the mosquito species involved in the transmission of the haemosporidian parasites Plasmodium spp. in two wild populations of breeding great tits (Parus major) in western Switzerland. Additionally, we compared Plasmodium lineages, based on mitochondrial DNA cytochrome b sequences, between the vertebrate and dipteran hosts, and evaluated the prevalence of the parasite in the mosquito populations. Plasmodium spp. were detected in Culexpipiens only, with an overall 6.6% prevalence. Among the six cytochrome b lineages of Plasmodium identified in the mosquitoes, three were also present in great tits. The results provide evidence for the first time that C. pipiens can act as a natural vector of avian malaria in Europe and yield baseline data for future research on the epidemiology of avian malaria in European countries. PMID:22506060

Avian malaria studies have taken a prominent place in different aspects of evolutionary ecology. Despite a recent interest in the role of vectors within the complex interaction system of the malaria parasite, they have largely been ignored in most epidemiological studies. Epidemiology of the disease is however strongly related to the vector's ecology and behaviour, and there is a need for basic investigations to obtain a better picture of the natural associations between Plasmodium lineages, vector species and bird hosts. The aim of the present study was to identify the mosquito species involved in the transmission of the haemosporidian parasites Plasmodium spp. in two wild populations of breeding great tits (Parus major) in western Switzerland. Additionally, we compared Plasmodium lineages, based on mitochondrial DNA cytochrome b sequences, between the vertebrate and dipteran hosts, and evaluated the prevalence of the parasite in the mosquito populations. Plasmodium spp. were detected in Culexpipiens only, with an overall 6.6% prevalence. Among the six cytochrome b lineages of Plasmodium identified in the mosquitoes, three were also present in great tits. The results provide evidence for the first time that C. pipiens can act as a natural vector of avian malaria in Europe and yield baseline data for future research on the epidemiology of avian malaria in European countries.

Background: The irritant effect of some insecticides can cause a proportion of mosquitoes to leave the sprayed rooms before acquiring a lethal dose, so the repeated contact al sub-lethal dose may lead to extent the resistance. Methods: Larvae and pupae of Culexpipiens complex were collected in mass from open canals of waste water in capital city Tehran and reared to obtain the first generation at laboratory. Sugar-fed 2–3 days female mosquitoes were used for the experiments and compared with laboratory strain. The irritability tests of insecticides impregnated papers were measured in plastic conical exposure chambers placed which implemented at controlled conditions according to the method described by WHO. Number of take-offs were counted during 15 minutes of exposure time. Results: DDT had the most irritancy effect against field population of Cx. pipiens. DDT, permethrin and deltamethrin was moderately irritable against laboratory strain, whereas, addition to three previous insecticides, malathion, cyfluthrin and propoxur should be also considered as moderately irritable insecticides for field population of. Irritability level of etofenprox, fenithrothion, bendiocarb, and lambdacyhalothrin did not differ from control group. Conclusion: The irritability response of mosquitoes may have a negative impact on control measures. Periodical execution of irritability tests with insecticides that routinely used in vector control program is highly recommended. PMID:27308276

The short day lengths of late summer are used to program the overwintering adult diapause (dormancy) of the Northern house mosquito, Culexpipiens. Here, we investigated the role of clock genes in initiating this diapause and asked whether the circadian cycling of clock gene expression persists during diapause. We provide evidence that the major circadian clock genes continue to cycle throughout diapause and after diapause has been terminated. RNA interference (RNAi) was used to knock down the core circadian clock genes and to then assess the impact of the various clock genes on the ability of females to enter diapause. RNAi directed against negative circadian regulators (period, timeless and cryptochrome2) caused females that were reared under diapause-inducing, short day conditions to avert diapause. In contrast, knocking down the circadian-associated gene pigment dispersing factor caused females that were reared under diapause-averting, long day conditions to enter a diapause-like state. Our results implicate the circadian clock in the initiation of diapause in C. pipiens. PMID:25653422

The residual effectiveness of VectoMax® WSP (a water-soluble pouch formulation containing a combination of Bacillus thuringiensis subsp. israelensis strain AM65-52 and B. sphaericus strain ABTS 1743) when applied to septic tanks against 3rd- and 4th-stage larvae of Culexpipiens L. was evaluated in this study. This formulation was evaluated at operational application rates of 1 pouch (10 g) and 2 pouches (20 g) per septic tank. Both application rates resulted in >96% control of larvae for 24 days. Operationally, VectoMax WSP has proven to be a useful tool for the nonchemical control of Culex species in septic tank environments.

To evaluate the vectorial capacity of mosquitoes for viruses in Japan, the host-feeding habits of the mosquitoes were analyzed by sequencing polymerase chain reaction-amplified fragments of the cytochrome b and 16S ribosomal RNA regions of the mitochondrial DNA of 516 mosquitoes of 15 species from seven genera that were collected from residential areas during 2003-2006. Culexpipiens L. and Aedes albopictus Skuse were the most commonly collected species in urban and suburban residential areas. Anautogenous Culexpipiens pallens Coquillett was distinguished from the autogenous Cx. pipiens form molestus Forskal using a polymerase chain reaction-based identification method. Both Cx. p. pallens and Cx. p. form molestus exhibited similar host-feeding habits, broadly preferring avian (50.0 and 42.5% of avian, respectively) and mammalian (38.6 and 45.0% of avian, respectively) hosts, such as tree sparrows, ducks, and humans. Conversely, Ae. albopictus exhibited a highly mammalophilic and anthropophilic feeding pattern, with 84.2% feeding on mammalian hosts and 68.5% of these on humans. We concluded that in Japan, Cx. pipiens might play a significant role in the avian-to-mammal transmission of viruses, such as West Nile virus, whereas Ae. albopictus might play a role in the human-human transmission of dengue and Chikungunya viruses.

Background: The aim of the current study was to develop a marking technique as an internal marker to mark post blood meal mosquitoes by using stable phosphate isotope 32P and determine the optimal concentration of it. Methods: An isotonic physiological saline solution, containing different concentration of radioactive isotope 32P-labeled disodium phosphate (Na2H32PO4) was injected into rabbits via the jugular vein in the laboratory. Emerged Cx. pipiens were marked after feeding on rabbit. At the same time, the labeled conditions of emerged Cx. pipiens were also measured by placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside. Results: According to the label condition of Cx. pipiens after taking blood and the effect of different dosage Na2H32PO4 on rabbit health, the optimal concentration of radioactive isotope was determined, that is, 0.1211 mCi/kg. By placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside, the emerged mosquitoes were also labeled. Therefore, feeding mosquitoes on the animal injected with radioactive Na2H32PO4 was more practical for detecting and tracing mosquitoes. Conclusion: The method was less time-consuming, more sensitive and safer. This marking method will facilitate post-bloodmeal studies of mosquitoes and other blood-sucking insects. PMID:27308279

We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culexpipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of F(ST) by 0.002. A 100-km increase in the longitudinal change caused an increased square root of F(ST) by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking.

We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culexpipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of FST by 0.002. A 100-km increase in the longitudinal change caused an increased square root of FST by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking. PMID:19861620

In this study, we identified blossoms that attract Culexpipiens L. s.l. in a Mediterranean habitat by using branches of 26 common plant species as baits for traps. The highest catch, 60.5% of the total, by flowers of Tamarix jordanis Boiss., was approximately 6 times greater than the 10.7% caught by flowering Polygonum equisetiforme Sm., and 10 times higher than the 6.6% caught by flowers of Acacia saligna (Lindle) H. L. Wendl. The catch elicited by the other plants ranged between 4.0 and 0.1%. Plant attraction also was evaluated in a field situation. Experimental and control sites were similar strips of vegetation along water channels with T. jordanis trees in the center. In the experimental site, these trees were sprayed with sucrose solution, food dye, and oral insecticide (Spinosad). Concurrently, patches of plant species and trees in the control site were sprayed with solutions of sucrose and different food dye markers. Cx. pipiens populations in both sites were monitored. The highest proportion (65.2%) of the marked mosquitoes in the control site carried the dye of flowering T. jordanis. The dye of flowering P. equisetiforme and that of A. saligna were found, respectively, in 8.1 and 3.5% of the labeled mosquitoes. The marker of reed groups (Phragmites australis [Cav.] Steudel) above the water was found in 19.4% of mosquitoes, whereas the different marker of dry land reeds was found in only 0.4% of the labeled mosquitoes. In the experimental site, after treatment, the mosquitoes decreased from approximately 255 per trap to approximately 24 mosquitoes per trap, whereas the catch in the control site reached approximately 400 mosquitoes per trap.

The larvicidal activity of hydrodistillate extracts from Chrysanthemum coronarium L., Hypericum scabrum L., Pistacia terebinthus L. subsp. palaestina (Boiss.) Engler, and Vitex agnus castus L. was investigated against the West Nile vector, Culexpipiens L. (Diptera: Culicidae). Yield and identification of the major essential oils from each distillation was determined by GC-MS analyses. The major essential oil component for each plant species was as follows: α-pinene for P. terebinthus palaestina, and H. scabrum (45.3% and 42.3%, respectively), trans-β-caryophyllene for V. agnus castus (22.1%), and borneol for C. coronarium (20.9%). A series of distillate concentrations from these plants (that ranged from 1 ppm to 500 ppm, depending on plant species) were assessed against late third to early fourth C. pipiens larvae at 1, 6, and 24 h posttreatment. In general, larval mortality to water treated with a distillate increased as concentration and exposure time increased. H. scabrum and P. terebinthus palaestina were most effective against the mosquito larvae and both produced 100% mortality at 250 ppm at 24-h continuous exposure compared with the other plant species. Larval toxicity of the distillates at 24 h (LC(50) from most toxic to less toxic) was as follows: P. terebinthus palaestina (59.2 ppm) > H. scabrum (82.2 ppm) > V. agnus castus (83.3 ppm) > C. coronarium (311.2 ppm). But when LC(90) values were compared, relative toxicity ranking changed as follows: H. scabrum (185.9 ppm) > V. agnus castus (220.7 ppm) > P. terebinthus palaestina (260.7 ppm) > C. coronarium (496.3 ppm). Extracts of native Turkish plants continue to provide a wealth of potential sources for biologically active agents that may be applied against arthropod pests of man and animals.

Essential or volatile oils of plants have been variously reported to have many medicinal applications. Methanol, acetone and petroleum ether extracts of Ocimum basilicum and Glycyrrhiza glabra were screened for their repellency effect against Culexpipiens mosquito. The repellent action of the present plants extracts were varied depending on the solvent used and dose of extract. Methanol extract of O. basilicum exhibited the lowest repellent activity as it recorded 77.4% at 6.7mg/cm2. The petroleum ether and acetone extract of 0. basilicum showed repellency of 98.1 & 84.6% respectively, at dose of 6.7mg/cm2, while methanolic extract of G. glabra recorded 73.8 & 50.3% at dose of 6.7 &1.7mg/cm2 respectively, the petroleum ether and acetone extract of G. glabra showed repellency of 76.3 & 81.6%, respectively at dose of 6.7mg/cm2, compared with the commercial formulation, N.N. diethyl toulamide (DEET) which exhibited 100% repellent action at dose of 1.8mg/cm2, respectively. The results may contribute to design an alternative way to control mosquitoes currently based on applications of synthetic insecticides. These extracts could be developed commercially as an effective personal protection meaure against mosquito bites and thus to control diseases caused by mosquito-borne pathogens.

The toxicity of pellitorine alone or in combination with (-)-asarinin, alpha-asarone, methyleugenol, or pentadecane (1:1, 1:2, 1:3, 2:1, and 3:1 ratios) to third instars from an insecticide-susceptible KS-CP strain and -resistant DJ-CP colony of Culexpipiens pallens Coquillett was evaluated using a direct-contact mortality bioassay. The binary mixture of pellitorine and (-)-asarinin (3:1 ratio) was significantly more toxic against KS-CP larvae (0.95 mg/liter) and DJ-CP larvae (1.07 mg/liter) than either pellitorine (2.08 mg/liter for KS-CP and 2.33 mg/liter for DJ-CP) or (-)-asarinin (11.45 and 12.61 mg/liter) alone. The toxicity of the other binary mixtures (1:1, 1:2, 1:3, and 2:1 ratios) and pellitorine did not differ significantly from each other. Based on the co-toxicity coefficient (CC) and synergistic factor (SF), the three binary mixtures (1:3, 2:1, and 3:1) operated synergistically (CC, 250-390 and SF, 1.4-2.2 for KS-CP; CC, 257-279 and SF, 1.1-2.1 for DJ-CP). The binary mixtures of pellitorine and (-)-asarinin merit further study as potential larvicides for the control of insecticide-resistant mosquito populations.

Adult females of the mosquito Culexpipiens entering diapause increase sugar water ingestion and reduce evaporative water loss, but how these attributes of the diapause program impact activity of the renal excretory system remains unknown. Here we compared the renal excretory capacity of diapausing and non-diapausing females, as well as the molecular expression of aquaporin (AQP) genes that encode channels involved in transporting water and/or small metabolites. Baseline urine excretion rates in diapausing mosquitoes were higher than in those of their non-diapausing counterparts, possibly a consequence of the intense sugar feeding associated with diapause. But, diapausing mosquitoes exhibited a much lower capacity for diuresis than non-diapausing mosquitoes. The suppressed diuretic capacity likely reflects reduced investment in the energetically-expensive post-prandial diuresis, an event not observed in diapausing mosquitoes. The mRNA expression levels of two genes encoding AQPs, Eglp1 and Aqp12L, in diapausing mosquitoes were down-regulated (on day 14) and up-regulated (on both days 3 and 14), respectively, in whole body samples. These changes were not evident in the excretory system (i.e., Malpighian tubules and hindgut), which showed no differential expression of AQPs as a function of diapause. Several AQP mRNAs were, however, differentially expressed in the midgut, ovaries, and abdominal body wall of diapausing mosquitoes, suggesting that AQPs in these tissues may be playing important non-excretory roles that are unique to diapause physiology.

The impact of weather variables on Culexpipiens L. (Diptera: Culicidae) population dynamics in the Po Valley, Northern Italy, a densely populated region containing the largest industrial and agricultural areas in Italy, was investigated. Monitoring of mosquitoes was carried out by using CO(2)-baited traps without light, collecting data weekly from 1700 to 0900 hours during the period May-September, from 1997 to 2011. Daily minimum, average, and maximum relative humidity; daily minimum, maximum, and average temperature; rainfall; and hydroclimatic balance (rainfall-potential evapotranspiration) were obtained from three weather stations within the surveillance zone. The average population dynamic trend over the 15-yr period showed a bell-shaped curve with a major peak in June and a secondary peak at the end of August in the rural areas, whereas bimodality was not evidenced in the urban areas. The correlation analyses showed that the mosquito seasonal population and the population in the period of maximum West Nile virus circulation (August-September) was mostly affected by the relative humidity registered from March to July, particularly in May, and, to a lower extent, also by hydroclimatic balance registered in April-July, and by the rainfall occurred in June-July. In addition, the rate of increase of the population during the spring months influenced the development of the mosquito population of the following months.

Excessive and continuous application of deltamethrin has resulted in the development of deltamethrin resistance among mosquitoes, which becomes a major obstacle for mosquito control. In a previous study, differentially expressed miRNAs between deltamethrin-susceptible (DS) strain and deltamethrin-resistant (DR) strain using illumina sequencing in Culexpipiens pallens were identified. In this study, we applied RNAi and the Centers for Disease Control and Prevention (CDC) bottle bioassay to investigate the relationship between miR-2∼13∼71 cluster (miR-2, miR-13 and miR-71) and deltamethrin resistance. We used quantitative real-time PCR (qRT-PCR) to measure expression levels of miR-2∼13∼71 clusters. MiR-2∼13∼71 cluster was down regulated in adult female mosquitoes from the DR strain and played important roles in deltamethrin resistance through regulating target genes, CYP9J35 and CYP325BG3. Knocking down CYP9J35 and CYP325BG3 resulted in decreased mortality of DR mosquitoes. This study provides the first evidence that miRNA clusters are associated with deltamethrin resistance in mosquitoes. Moreover, we investigated the regulatory networks formed between miR-2∼13∼71 cluster and its target genes, which provide a better understanding of the mechanism involved in deltamethrin resistance.

The interaction of the mosquito and the invading virus is complex and can result in physiological and gene expression alterations in the insect. The association of West Nile virus (WNV) and Culexpipiens quinquefasciatus mosquitoes results in measurable changes in gene expression; 22 gene products were shown previously to have altered expression. Sequence analysis of one product, CQ G1A1, revealed 100% amino acid identity to gram negative bacteria binding proteins (CPQGBP) in Cx. p. quinquefasciatus, Aedes aegypti (70%) and Anopheles gambiae (63%) that function in pathogen recognition. CQ G1A1 also was differentially expressed following WNV infection in two populations of Cx. p. quinquefasciatus colonized from Florida with known differences in vector competence for WNV and showed spatial and temporal gene expression differences in midgut, thorax, and carcass tissues. These data suggest gene expression of CQ G1A1 is influenced by WNV infection and the WNV infection-controlled expression differs between populations and tissues. PMID:27999244

Within developing ovaries of many insects, some developing follicles or oocytes usually degenerate (follicular atresia or oosorption), while the others may continue to grow to maturity, thus maintaining the balance between the number of eggs and reproductive circumstances such as available nutrients. To help clarify the phenomenon of follicular atresia during ovarian development, we examined cysteine proteinases stored in mosquito Culexpipiens pallens ovaries. First, analysis using synthesized substrates showed that cathepsin B- and L-like proteinases gradually accumulated in the developing ovaries after a blood meal, which required more than 10 min of preincubation under acidic conditions to reach their maximum activities. However, homogenates of degenerating follicles 3 days after feeding showed proteolytic activities without acid treatment, suggesting that the proteinases had already been activated, while the extract of normally developing follicles collected from the same ovaries required more than 10 min of acid preincubation to reach the optimum activities, suggesting that the enzymes remained as inactive forms. Chemical and immunohistochemical analyses showed that more proteinases are located in the cytoplasm, rather than being associated with yolk granules. Ovarian proteinases, which are believed to become activated at the onset of embryogenesis, should also be activated during oogenesis, presumably to enhance oosorption.

The larvicidal activity of chlorpyrifos-methyl and temephos was evaluated against Culexpipiens L. (Diptera: Culicidae) in septic tanks in Antalya, Turkey. Chlorpyrifos-methyl (Pyrifos MT 25 emulsifiable concentrate [EC] ) was evaluated at application rates of 0.04, 0.08, and 0.12 mg active ingredient (AI)/liter, and temephos (Temeguard 50 EC) was evaluated at 0.02, 0.04, and 0.06 mg (AI)/liter during a 21-d study. Generally, overall larval reduction in septic tanks from single- and multifamily dwellings treated with either larvicide was significantly greater than pretreatment levels and control tanks for the duration of the study. At 14 d posttreatment, duration of control was greatest in multifamily tanks treated with chlorpyrifos-methyl at the highest application rate with similar levels of control through 21 d for single-family dwellings (range 97-100%). Septic tanks from both types of family dwellings treated at the highest application rate of temephos resulted in >90% reduction through day 21 (range 91-100%). Laboratory bioassays of septic tank water treated at field application rates, without daily dilution, revealed that complete larval mortality was achieved for 21 d at each application rate and formulation. It is thought that daily addition of water and organic matter to the septic tanks in the single and multifamily dwellings influenced the duration of effectiveness of the larvicides.

We studied effects of combined diethylcarbamazine (DEC) and albendazole (ALB) treatment on Wuchereria bancrofti microfilaria (MF) uptake and development of infective larvae (L3) in Culexpipiens. Consenting Egyptian adults with microfilaremia (MF > 300/mL) were treated with one or seven daily doses of DEC/ALB. Laboratory-reared mosquitoes were fed on subjects before and after treatment. MF uptake and infectivity (assessed by mosquito dissection) were reduced by 89.6% and 82.9%, respectively, 12 months after single-dose treatment and by 96.2% and 99.7%, respectively, after multi-dose treatment. The L3:mosquito ratio decreased by 88% to 0.082 after single-dose treatment and by 99.8% to 0.001 after multi-dose treatment. If high coverage rates can be achieved for several annual cycles, mass drug administration (MDA) with DEC/ALB has the potential to decrease transmission to unsustainable levels and eliminate filariasis in populations. Multi-dose MDA (especially in the first year) might interrupt transmission with fewer cycles than single-dose treatment.

Fecundity reduction is a well-recognized phenomenon of parasite infection in insects. Reduced production of eggs might increase longevity of a host and release nutrients to both host and parasite that would otherwise be used for oogenesis. The objective of this study was to assess effects on fecundity caused by Hepatozoon sipedon, an apicomplexan blood parasite of snakes, in its invertebrate host, the mosquito Culexpipiens. In the first gonotrophic cycle, the mean number of eggs laid by mosquitoes infected with H. sipedon did not differ significantly from those laid by uninfected mosquitoes. However, in the second gonotrophic cycle infected mosquitoes laid significantly fewer eggs than did uninfected mosquitoes, and fecundity was reduced by 100% in mosquitoes with parasite burdens of more than 60 oocysts. There was a significant negative correlation between parasite burden, or the number of oocysts, and the number of eggs produced in the second gonotrophic cycle. Significantly fewer viable larvae hatched from eggs laid by infected compared to uninfected mosquitoes in the second gonotrophic cycle. These data indicate that fecundity reduction occurs in this system, although the physiological mechanisms driving this phenotype are not yet known.

High rates of West Nile virus (WNV) transmission to humans are associated with exceptionally hot and dry summers. This is paradoxical since the eggs of Culex vectors of WNV depend on the persistence of containers with water, which decline during droughts. We examined the effects of forced-egg retention on the reproductive success of female Culexpipiens as well as behavioral responses, such as likelihood of secondary blood meals. As controls we examined the effects of female age and delayed mating. We found that early mating is essential to achieve reproductive success and, consistent with an "all-or-none" ovipositing strategy, C. pipiens females are able to retain considerable reproductive potential while searching for oviposition sites. Specifically, although forced-egg retention resulted in significant decreases in fitness, the decline was moderate for 5 weeks and most can be accounted for by increases in female age. Consequently, no females took blood more than once per gonotrophic cycle, which eliminates the possibility that heightened vectorial capacity due to multiple blood-feedings increases WNV transmission during periods of drought. Instead, our findings suggest that during droughts populations of C. pipiens have time to locate the remaining water holes, which are associated with human populations and WNV-competent bird species.

The genome of all densoviruses (DNVs) so far isolated from mosquitoes or mosquito cell lines consists of a 4-kb single-stranded DNA molecule with a monosense organization (genus Brevidensovirus, subfamily Densovirinae). We previously reported the isolation of a Culexpipiens DNV (CpDNV) that differs significantly from brevidensoviruses by (i) having a approximately 6-kb genome, (ii) lacking sequence homology, and (iii) lacking antigenic cross-reactivity with Brevidensovirus capsid polypeptides. We report here the sequence organization and transcription map of this virus. The cloned genome of CpDNV is 5,759 nucleotides (nt) long, and it possesses an inverted terminal repeat (ITR) of 285 nt and an ambisense organization of its genes. The nonstructural (NS) proteins NS-1, NS-2, and NS-3 are located in the 5' half of one strand and are organized into five open reading frames (ORFs) due to the split of both NS-1 and NS-2 into two ORFs. The ORF encoding capsid polypeptides is located in the 5' half of the complementary strand. The expression of NS proteins is controlled by two promoters, P7 and P17, driving the transcription of a 2.4-kb mRNA encoding NS-3 and of a 1.8-kb mRNA encoding NS-1 and NS-2, respectively. The two NS mRNAs species are spliced off a 53-nt sequence. Capsid proteins are translated from an unspliced 2.3-kb mRNA driven by the P88 promoter. CpDNV thus appears as a new type of mosquito DNV, and based on the overall organization and expression modalities of its genome, it may represent the prototype of a new genus of DNV.

Background Culex is an important mosquito as vectors for the transmission of serious diseases, such as filariasis, West Nile virus, dengue, yellow fever, chikungunya and other encephalitides. Nearly one billion people in the developing countries are at risk. In order to discover new bioactive molecules and pesticides acting on mosquito, we designed active amide structure and synthesized a series of novel diamide derivatives. Results A series of novel diamide derivatives were designed and synthesized. Their structures were characterized by 1 H NMR, FTIR and HRMS. The single crystal structure of compound 6n was determined to further elucidate the structure. Biological activities of these compounds were tested. Most of them exhibited higher mosquito larvicidal activity. Especially compound 6r displayed relatively good activity to reach 70% at 2 μg/mL. Conclusion A practical synthetic route to amide derivatives by the reaction of amide with another acid is presented. This study suggests that the diamide derivatives exhibited good effective against mosquito. PMID:22963735

Host-seeking heights, host-seeking activity patterns, and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culexpipiens complex from July to December 2002, by using chicken-baited can traps (CT) at four ecologically different sites in Shelby County, TN. Host-seeking height was assessed by CT placed at elevations of 3.1, 4.6, and 7.6 m during one 24-h period per month. Host-seeking activity was assessed by paired CT placed at an elevation of 4.6 m. Can traps were sampled at one 10-h daytime interval and at seven 2-h intervals during the evening, night, and morning. Cx. pipiens complex mosquitoes accounted for 87.1% of collected mosquitoes. Culex (Melanoconion) erraticus (Dyar & Knab) accounted for 11.9% of specimens. The average number of Cx. pipiens complex mosquitoes collected per 24-h CT period from July to September was lowest at a rural middle income site (1.7), intermediate at an urban middle income site (11.3), and highest at an urban low income site (47.4). Can traps at the forested site failed to collect Cx. pipiens complex mosquitoes. From July to September at urban sites, Culexpipienspipiens L. was the rarest of the three complex members accounting for 11.1-25.6% of specimens. At the rural site, Culexpipiens quinquefasciatus Say was the rarest member of the complex. Cx. p. pipiens was not collected after September. Mean abundance of Cx. pipiens complex mosquitoes was higher in traps at 7.6 m than in traps at 4.6 m. Abundances at 3.1 m were intermediate and not significantly different from abundances at the other heights. Initiation of host-seeking activity was associated with the end of civil twilight and activity occurred over an extended nighttime period lasting 8-10 h. All 11 WNV-positive mosquitoes were Cx. pipiens complex mosquitoes collected from urban sites in traps placed at elevations of 4.6 and 7.6 m. Infection rates were marginally nonsignificant by height. Infection rates, host

In an effort to determine whether female Culexpipiens L. and Culex restuans Theobald mosquitoes (Diptera: Culicidae) are attracted to crow, Corvus brachyrhynchus, uropygial gland secretions, CDC miniature light traps (baited with CO2 but with the lights removed) were placed at approximately 1.5- and 5-m elevations, in 10 trees in awoodlot near Niagara Falls, Canada. These traps were assigned either a bird odor or a blank control. Bird odors were created by attaching cotton swabs coated with crow uropygial gland secretions to the trap intake. A significantly greater number of Cx. pipiens/ restuans were found in the 5-m traps compared with 1.5-m traps, with a significant number attracted to the bird odor over the no odor traps at the 5-m elevation, but not at 1.5 m. We also found more Aedes vexans (Meigen) in the 1.5-m traps than the 5-m traps; however, presence or absence of bird odor did not influence the distribution of Ae. vexans.

The Horn of Africa represents a region formerly known to be highly susceptible to mosquito-borne infectious diseases. In order to monitor and analyze the current presence and threat of vector mosquitoes, continuous and standardized trapping using CDC light traps without an additional CO2-generator has been carried out at six selected monitoring sites located in Djibouti City, from August 2008 until December 2009. An overall of 620 haematophageous Diptera were trapped, 603 (97.3%) were mosquitoes, 10 (1.6%) were sand flies, and 7 (1.1%) were biting midges, respectively. Genus distribution of mosquitoes revealed that 600 (99.5%) were Culex spp., 2 (0.3%) were Anopheles sergentii, and 1 (0.2%) was Aedes aegypti. Culex species were represented by Cx. quinquefasciatus (78.5%), and Cx. pipiens ssp. torridus (21.5%). The later species was first detected focally in early December 2009 showing a strongly increasing population density resulting in a maximum trap rate of 25 mosquitoes per trap night. Sand flies were all Sergentomyia antennata, and biting midges of the genus Culicoides were represented by C. nubeculosus (71.4%) and C. vexans (28.6 %). The findings included the first records for Cx. pipiens ssp. torridus and An. sergentii in Djibouti. However, none of the captured female Culex spp, the known vector for West Nile Virus, showed positive results for viral nucleic acids using WNV RT-real time PCR system. Also, females An. sergentii were Plasmodium falciparum and P. vivax circumsporozoite protein negative.

Background The recent spread of West Nile Virus in temperate countries has raised concern. Predicting the likelihood of transmission is crucial to ascertain the threat to Public and Veterinary Health. However, accurate models of West Nile Virus (WNV) expansion in Europe may be hampered by limited understanding of the population dynamics of their primary mosquito vectors and their response to environmental changes. Methods We used data collected in north-eastern Italy (2009–2011) to analyze the determinants of the population growth rate of the primary WNV vector Culexpipiens. A series of alternative growth models were fitted to longitudinal data on mosquito abundance to evaluate the strength of evidence for regulation by intrinsic density-dependent and/or extrinsic environmental factors. Model-averaging algorithms were then used to estimate the relative importance of intrinsic and extrinsic variables in describing the variations of per-capita growth rates. Results Results indicate a much greater contribution of density-dependence in regulating vector population growth rates than of any environmental factor on its own. Analysis of an average model of Cx. pipiens growth revealed that the most significant predictors of their population dynamics was the length of daylight, estimated population size and temperature conditions in the 15 day period prior to sampling. Other extrinsic variables (including measures of precipitation, number of rainy days, and humidity) had only a minor influence on Cx. pipiens growth rates. Conclusions These results indicate the need to incorporate density dependence in combination with key environmental factors for robust prediction of Cx. pipiens population expansion and WNV transmission risk. We hypothesize that detailed analysis of the determinants of mosquito vector growth rate as conducted here can help identify when and where an increase in vector population size and associated WNV transmission risk should be expected. PMID:24428887

Background The Middle East North Africa (MENA) region is under continuous threat of the re-emergence of West Nile virus (WNV) and Rift Valley Fever virus (RVF), two pathogens transmitted by the vector species Culexpipiens. Predicting areas at high risk for disease transmission requires an accurate model of vector distribution, however, most Cx. pipiens distribution modeling has been confined to temperate, forested habitats. Modeling species distributions across a heterogeneous landscape structure requires a flexible modeling method to capture variation in mosquito response to predictors as well as occurrence data points taken from a sufficient range of habitat types. Methods We used presence-only data from Egypt and Lebanon to model the population distribution of Cx. pipiens across a portion of the MENA that also encompasses Jordan, Syria, and Israel. Models were created with a set of environmental predictors including bioclimatic data, human population density, hydrological data, and vegetation indices, and built using maximum entropy (Maxent) and boosted regression tree (BRT) methods. Models were created with and without the inclusion of human population density. Results Predictions of Maxent and BRT models were strongly correlated in habitats with high probability of occurrence (Pearson’s r = 0.774, r = 0.734), and more moderately correlated when predicting into regions that exceeded the range of the training data (r = 0.666,r = 0.558). All models agreed in predicting high probability of occupancy around major urban areas, along the banks of the Nile, the valleys of Israel, Lebanon, and Jordan, and southwestern Saudi Arabia. The most powerful predictors of Cx. pipiens habitat were human population density (60.6% Maxent models, 34.9% BRT models) and the seasonality of the enhanced vegetation index (EVI) (44.7% Maxent, 16.3% BRT). Maxent models tended to be dominated by a single predictor. Areas of high probability corresponded with sites of

The aim of this study was to assess whether certain attributes of larval breeding sites are correlated with pupal productivity (i.e. numbers of pupae collected per sampling period), so that these could be used as the focus for control measures to enhance control efficiency. Therefore, the objectives were to identify the months of highest pupal productivity of Aedes aegypti (L.) and Culexpipiens L. (Diptera: Culicidae) in an urban temperate cemetery in Argentina where artificial containers of < 6 L (flower vases) were the predominant breeding habitats, to compare various measures of the productivity of sunlit and shaded containers and to determine whether the composition of the containers affected pupal productivity. Over a period of 9 months, 200 randomly chosen water-filled containers (100 sunlit and 100 shaded), out of approximately 3738 containers present (approximately 54% in shade), were examined each month within a cemetery (5 ha) in Buenos Aires (October 2006 to June 2007). In total, 3440 immatures of Cx pipiens and 1974 of Ae. aegypti were collected. The larvae : pupae ratio was 10 times greater for the former, indicating that larval mortality was greater for Cx pipiens. Both mosquito species showed a higher container index (CI) in shaded than in sunlit containers (Ae. aegypti: 12.8% vs. 6.9% [chi(2) = 17.6, P < 0.001]; Cx pipiens: 6.3% vs. 1.8% [chi(2) = 24, P < 0.001]). However, the number and the density of immatures per infested container and the number of pupae per pupa-positive container did not differ significantly between sunlit and shaded containers for either species. Therefore, the overall relative productivity of pupae per ha of Ae. aegypti and Cx pipiens was 2.3 and 1.8 times greater, respectively, in shaded than in sunlit areas as a result of the greater CIs of containers in shaded areas. Neither the CI nor the number of immatures per infested container differed significantly among container types of different materials in either lighting

Salivary glands of Culexpipiens and Aedes caspius were analyzed to determine total protein content and its fractionation during adult female development and after blood sucking. In both species, the molecular weight of proteins ranged between 26.000 and 84.000 Daltons. These proteins were not identical in the two species. In Cx. pipiens, the total protein level increased during the first 3 days of adult development from 4.94 +/- 0.84 to 6.6 +/- 0.37 micrograms/gland. During this period, the salivary gland proteins were separated into 35, 34 and 37 fractions respectively. Cx. pipiens released in the human host 64% of the total proteins while taking a blood meal compared to unfed females. This decrease in protein level was proportional to protein fractions. Over the next 6 days, the protein level increased again to attain values comparable to those obtained prior to blood sucking. In Ae. caspius, the total protein level of the salivary glands did not change during the first 4 days of adult development (range between 3.13 +/- 0.27 and 3.91 +/- 0.36 micrograms gland), but on the fifth day, 2-fold increase was observed. The total salivary gland protein increased during the next 3 days after blood sucking to reach 15.5 +/- 0.98 micrograms/gland. During this period, a tremendous change in protein patterns was observed. After oviposition, on the fourth day, a significant reduction in the total protein level was observed (4.13 +/- 0.56 micrograms/gland), but over the next 3 days the level increased again (range between 4.13 +/- 0.66 and 7.13 +/- 0.66 micrograms/gland).

Culexpipiens (Diptera: Culicidae) is the most widely distributed mosquito species in Algeria and many other countries in the world. Mosquitoes are generally controlled by conventional insecticides but these may pose strong secondary effects on the environment. In this context, the insect growth regulators (IGRs) have shown promise in controlling pest insects. Halofenozide (23% EC) is a novel IGRs belonging to the class of non-steroidal ecdysone agonists, and it was found toxic for larvae of C. pipiens. In addition biological methods constitute an alternative to chemical control. Several fish species have been tested against mosquitoes, and Gambusia affinis was found very efficient. In the present study we evaluated the impact of this new potent insecticide (halofenozide) on growth and metric indexes in the larvivorous fish G. affinis under laboratory conditions. In addition, the effects were evaluated on the enzymatic activities of acetyl cholinesterase (AChE) and glutathione S-transferase (GST). The insecticide was added in water at two concentrations (12.6 and 28.6 microg/L) corresponding to the LC50 and LC90 obtained against fourth instar larvae of C. pipiens, and adult females of G. affinis were exposed to halofenozide for 30 days. At different exposure times we measured the length and weight of fishes, the index of condition (K), the gonado-somatic ratio (GSR) and the hepato-somatic ratio (HSR). The results showed that halofenozide had no significant (p>0.05) effects on growth, metric indexes and AChE activities. However, treatment caused a significant induction (p<0.05) in GST activities at days 15 and 30 with the highest dose. Our results indicate that this ecdysteroid agonist presented only minor secondary effects on the non-target fish species, and so it has potential for controlling of mosquitoes in an integrated manner.

The chemical composition of 14 essential oils (EOs), obtained from various parts (leaves, fruits, wood) of the six indigenous in Greece Juniperus family taxa, was determined by GC and GC/MS analysis. The insecticidal properties of these EOs were evaluated against Culexpipiens L. larvae of 3rd and early 4th instars, in order to delineate the relationship between the phytochemical content of the EOs and their larvicidal activities. The analytical data indicated that the EOs mainly consisted of monoterpenes, mostly cyclic and only occasionally aliphatic, and to a lesser percent, of diterpenes. The larvicidal bioassays against C. pipiens larvae revealed that the most active EO was derived from the wood of Juniperus drupacea and contains mainly non-oxygenated monoterpenes and a significant amount of diterpenes, displaying the highest chemodiversity. Its initial LC(50) value was 26.47 mg L(-1). On the contrary, the EO isolated from J. phoenicea berries, which consisted of monoterpenes (non-oxygenated, cyclic), was the less active displaying an LC(50) value of 96.69 mg L(-1). In respect to the contained phytochemicals, myrcene was assayed as the most toxic, displaying an LC(50) value of 33.83 mg L(-1), while the four isomers of pinene abundant in all EOs were less active exhibiting LC(50) values ranging from 70.40 to 94.88 mg L(-1). Results herein reveal that the EOs isolated from the studied Juniperus family taxa represent an inexpensive source of natural mosquito control mixtures.

Culexpipiens mosquito is a species widely spread across Europe and represents a competent vector for many arboviruses such as West Nile virus (WNV), which has been recently circulating in many European countries, causing hundreds of human cases. In order to identify the main determinants of the high heterogeneity in Cx. pipiens abundance observed in Piedmont region (Northwestern Italy) among different seasons, we developed a density-dependent stochastic model that takes explicitly into account the role played by temperature, which affects both developmental and mortality rates of different life stages. The model was calibrated with a Markov chain Monte Carlo approach exploring the likelihood of recorded capture data gathered in the study area from 2000 to 2011; in this way, we disentangled the role played by different seasonal eco-climatic factors in shaping the vector abundance. Illustrative simulations have been performed to forecast likely changes if temperature or density–dependent inputs would change. Our analysis suggests that inter-seasonal differences in the mosquito dynamics are largely driven by different temporal patterns of temperature and seasonal-specific larval carrying capacities. Specifically, high temperatures during early spring hasten the onset of the breeding season and increase population abundance in that period, while, high temperatures during the summer can decrease population size by increasing adult mortality. Higher densities of adult mosquitoes are associated with higher larval carrying capacities, which are positively correlated with spring precipitations. Finally, an increase in larval carrying capacity is expected to proportionally increase adult mosquito abundance. PMID:27105065

Laboratory observations on the effect of Hepatozoon gracilis on the egg production of the mosquito Cx. (Cx.) pipiens Linneaus under laboratory conditions revealed that H. gracilis infected mosquitoes produced significantly fewer eggs than uninfected ones. The egg production decreased as parasite burdens increased. Reduction in blood meal size in infected females did not reduce fecundity. No size differences was detected between oocyst-infected and uninfected females although sporozoite positive females were significantly large. Preoviposition period was affected significantly, while incubation period and percentage of egg hatching showed no significant changes. The longevity of female infected mosquitoes decreased insignificantly than in uninfected ones.

The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culexpipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands.

Background: The present study was undertaken to study the larvicidal activity of different extracts of Nerium oleander leaves, and post-treatment temperature- toxicity relationship of these extracts against Culexpipiens. Further, the most potent extract was used to evaluate its biological and genotoxic activities. Methods: Crude extracts of N. oleander leaves were prepared using water, chloroform, acetone and diethyl ether as solvents. Extraction was carried out using soxhlet apparatus. Bioassay test was carried out on the larvae, and the LC50 of each extract was determined. Thus, newly hatched first instar larvae were treated, and the mortality count was recorded daily till pupation (accumulated mortality). The LC50 of diethyl ether extract, as the most potent extract, was used for the further biological and genotoxic studies. Results: The results obtained indicated that diethyl ether extract of N. oleander leaves was the most potent extract, with LC50 of 10500 mg/l. The toxicity of the four extracts, using the LC50, at 10 °C was higher than that at 35 °C. The LC50 of diethyl ether extract significantly decreased the larval duration, pupal duration, percentage of pupation, percentage of adult emergence, longevity of females, fecundity, and oviposition activity index, whereas the growth index and the percentage of development per day of larvae and pupae were significantly increased compared to non-treated insects. Moreover, treatment with this extract induced significant dominant lethality in both male and female adults. Conclusion: It appears that diethyl ether extract of N. oleander leaves is potential control agent to Cx. pipiens. PMID:27047967

Background West Nile Virus (WNV) is an emerging global health threat. Transmission risk is strongly related to the abundance of mosquito vectors, typically Culexpipiens in Europe. Early-warning predictors of mosquito population dynamics would therefore help guide entomological surveillance and thereby facilitate early warnings of transmission risk. Methods We analysed an 11-year time series (2001 to 2011) of Cx. pipiens mosquito captures from the Piedmont region of north-western Italy to determine the principal drivers of mosquito population dynamics. Linear mixed models were implemented to examine the relationship between Cx. pipiens population dynamics and environmental predictors including temperature, precipitation, Normalized Difference Water Index (NDWI) and the proximity of mosquito traps to urban areas and rice fields. Results Warm temperatures early in the year were associated with an earlier start to the mosquito season and increased season length, and later in the year, with decreased abundance. Early precipitation delayed the start and shortened the length of the mosquito season, but increased total abundance. Conversely, precipitation later in the year was associated with a longer season. Finally, higher NDWI early in the year was associated with an earlier start to the season and increased season length, but was not associated with abundance. Proximity to rice fields predicted higher total abundance when included in some models, but was not a significant predictor of phenology. Proximity to urban areas was not a significant predictor in any of our models. Predicted variations in start of the season and season length ranged from one to three weeks, across the measured range of variables. Predicted mosquito abundance was highly variable, with numbers in excess of 1000 per trap per year when late season temperatures were low (average 21°C) to only 150 when late season temperatures were high (average 30°C). Conclusions Climate data collected early in

Two actin genes cloned from Culexpipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized actin that was most pronounced in the midguts of diapausing mosquitoes that were exposed to low temperature. In nondiapausing mosquitoes reared at 25°C and in diapausing mosquitoes reared at 18°C, polymerized actin was clustered at high concentrations at the intersections of the muscle fibers that form the midgut musculature. When adults 7–10 days post-eclosion were exposed to low temperature (-5°C for 12h), the polymerized actin was evenly distributed along the muscle fibers in both nondiapausing and diapausing mosquitoes. Exposure of older adults (1month post-eclosion) to low temperature (−5°C for 12h) elicited an even greater distribution of polymerized actin, an effect that was especially pronounced in diapausing mosquitoes. These changes in gene expression and actin distribution suggest a role for actins in enhancing survival of diapausing adults during the low temperatures of winter by fortification of the cytoskeleton. PMID:17078965

Climate change and West Nile fever (WNV) are both subjects of global importance. Many mosquitoes and the diseases they carry, including West Nile virus (WNV), are sensitive to temperature increase. The current study analyzes the lag correlations between weather conditions (especially air temperature) and 1) Culexpipiens mosquito population abundance, and 2) WNF frequency in humans, between 2001 and 2005 in Israel. These 5 years follow a long period with a documented tendency for temperature increase in the hot season in the country. Monthly anomalies of minimum and maximum temperatures, relative seasonal rainfall contribution, mosquito samplings (hazard level), and WNF cases (hospital admission dates and patients' addresses) were analyzed. Logistic regression was calculated between the climatic data and the mosquito samples, as Spearman correlations and Pearson cross-correlations were calculated between daily temperature values (or daily precipitation amounts) and the hospital admission dates. It was found that the disease appearance reflects the population distribution, while the risk tends to escalate around the metropolis characterized by an urban heat island. Positive anomalies of the temperature during the study period appear to have facilitated the mosquito abundance and, consequently, the disease emergence in humans. An important finding is the potential influence of extreme heat in the early spring on the vector population increase and on the disease's appearance weeks later. Awareness of such situations at the beginning of the spring may help authorities to reduce the disease risk before it becomes a real danger.

The North Shore Mosquito Abatement District applies extended release larvicides including methoprene-based Altosid® XR Extended Residual Briquets to approximately 40,000 catch basins in the southern half of the District's operational area at the beginning of each season. Treatments begin in May and typically again 9 to 10 wk later when larvicide efficacy appears to wane. In 2013 spinosad-based Natular™ XRT tablets were applied to basins, and a subset were monitored for larvae and pupae weekly with a standard dipper. When setting the threshold for retreatment as 12 juveniles per dip sample it was observed that basins required a second application 9 wk after the initial application, a time period similar to Altosid despite utilizing a different active ingredient. Average counts of weekly larval samples appeared to be positively associated with average numbers of Culexpipiens collected the following week in a gravid trap located among catch basins, highlighting the importance of basins as sources of these mosquitoes.

We cloned the gene that encodes prothoracicotropic hormone (PTTH) in the northern house mosquito, Culexpipiens, and investigated its expression profile in short-day (diapause-destined) and long-day (nondiapause-destined) individuals from the fourth-instar larval stage to 2 months of adulthood, as well as after a blood meal. The deduced C. pipiens PTTH (Cupip-PTTH) amino acid sequence contains seven cysteines with a specific spacing pattern. Sequence alignment suggests that Cupip-PTTH is 23% identical to Drosophila melanogaster PTTH, but is ≥59% identical to the PTTHs of other mosquitoes. Cupip-PTTH has structural characteristics similar to those of Bombyx mori PTTH and some vertebrate nerve growth factors with cysteine-knot motifs. PTTH transcripts exhibit a daily cycling profile during the final (fourth) larval instar, with peak abundance occurring late in the scotophase. The fourth-larval instar stage is one day longer in short-day larvae than in long-day larvae, resulting in larger larvae and adults. This additional day of larval development is associated with one extra PTTH cycle. No cycling was observed in pupae, but PTTH transcripts were slightly higher in short-day pupae than in long-day pupae throughout much of the pupal stage. PTTH expression persisted at a nearly constant level in diapausing adult females for the first month but then dropped by ∼50%, while expression decreased at the beginning of adulthood in nondiapausing females and then remained at a low level as long as the females were denied a blood meal. However, when nondiapausing females were offered a blood meal, PTTH transcripts rose approximately 7 fold in 2 h and remained elevated for 24 h. A few diapausing females (∼10%) will take a blood meal when placed in close proximity to a host, but much of the blood is ejected and such meals do not result in mature eggs. Yet, elevated PTTH mRNA expression was also observed in diapausing females that were force fed. Our results thus point to

A microsporidian, Nosema algerae Vavra and undeen, was found parasitizing larvae and adults of a laboratory colony of Culexpipiens L. originated from Gharbia Governorate. A detailed examination of the developmental stages of the pathogen under light and electron microscopes showed that they are typical of the original characteristics of N. algerae. Other observations on the infected individuals revealed that spores of the pathogen were found in all organs of the infected mosquitoes except the nervous system. The most heavily infected organ was the alimentary canal, particularly the mid gut. The susceptibility of the different larval instars of a laboratory maintained Cx. pipiens colony to infection by N. algerae was determined. Using 24 hours exposure to a range of doses between 40 and 5 x 10(5) spores per cm, larvae of the 1st and 2nd instars were more susceptible to infection than 3rd and 4th instar larvae as indicated by the differences in the estimated IC50 dosages. The dosages of N. algerae which produced 50% mortality (LC50) in each group of treated larvae was approximately 25-30 times higher than the doses that caused 50% infection. When a sublethal dose of 1.2 x 10(3) spores per cm2 of N. algerae was used to induce infection in each of the different larval instars infection rates of 100% were obtained in all exposed larval instars and in adults developed from them. Females from larvae infected with N. algerae had significantly reduced fecundity, fertility, and longevity regardless the larval instar in which infection was initiated. However, the reduction was obviously high when 1st and 2nd instars were exposed. The accumulative effects of reduced survival and fecundity on the reproductive potential of the infected females derived from larvae that were infected as 1st and 2nd instars probably serve to limit the natural increase of mosquito populations. Further studies are necessary, however, to determine the efficacy of this pathogen on target species in

The distribution of resting Culexpipiens s.l., L. in vegetation at the margins of breeding sites and the effects of a narrow, surrounding, sprayed belt of sugar, food dye and toxin on adult mosquitoes were studied near two pairs of control and experimental sewage ponds close to human habitation in the Judean hills. Control belts were without toxin. A sprayed belt of sugar and toxin 0.5 m from the water gradually reduced the population to an average of 38.3 mosquitoes per trap, 7.6% of the highest catch, which was 504.6 mosquitoes per trap in the control site. In the second experiment, in which bait belts were 5 m from the water, the toxic bait spraying was followed by a rise in catches from 207.9 to 274.9 mosquitoes. This was 41% of the 670.2 mosquitoes per trap in the parallel control site. In areas without toxin treatment, diurnal catches by net amounted to 20,705 mosquitoes. Of these, 86.1% (17,825) were caught within 1m of the water while only 8.2% (1701) were caught at a distance of 3 m. The remainders were caught up to 20 m away. Parity status was determined for female samples caught by net. In areas without toxin, parous females accounted for 37% of the catch and 13.2% were young, meconium containing specimens. The population diminished following spraying of toxic bait 1m from the water and included 13% parous females and 17.6% had meconium in the gut.

Culexpipiens molestus Forskal (Diptera: Culicidae) is the dominant mosquito species in septic tanks in South Korea. An assessment was made of the biological control potential of mud loaches, Misgurnus mizolepis Günther (Cypriniformes: Cobitidae), toward Cx. p. molestus larvae in laboratory and septic tanks. Results were compared with those of temephos 20% emulsifiable concentrate. In laboratory tests, all mud loaches survived on sedimentation chamber- and effluent chamber-collected water of aerobic septic tanks (ASTs), whereas all mud loaches died within 3-12 h after introduction into sedimentation chamber- and effluent chamber-collected water of anaerobic septic tanks, Gill hyperplasia and hemorrhages at the bases of pectoral fins were detected in all dead mud loaches. These appeared to have been caused by bacterial disease, rather than the physical and chemical characteristics of the septic tank water. A mud loach consumed an average range of 1,072-1,058 larvae of Cx. p. molestus in the AST water at 24 h. At the manufacturer's recommended rate (10 ml/ton) in the AST water, the temephos formulation did not cause fish mortality. In the AST experiment, predation of mosquito larvae by mud loaches at a release rate of one fish per 900 mosquito larvae resulted in complete mosquito control from the third day after treatment throughout the 18-wk survey period, compared with temephos 20% emulsifiable concentrate-treated AST water (reduction rate, 40% at 28 days after treatment). Reasonable mosquito control in aerobic septic tanks can be achieved by mosquito breeding season stocking of a rate of one mud loach per 900 mosquito larvae.

This work monitored changes in some digestive enzymes (trypsin and aminopeptidase) associated with the building up of resistance in Cx. pipiens larvae to two chemical insecticides (methomyl and/or malathion) and one biological insecticide (Bacillus thuringiensis-H14 or B.t H 14). The LC50 value of methomyl for both field- and the 12th generation (F12) of the selected strain was 1.789 ppm and 8.925 ppm respectively. The LC50 value of malathion for both field and the F12 of the selected strain was 0.082 ppm and 0.156 ppm respectively, and those of B.t H14 of field strain and the F12 was 2.550ppm & 2.395ppm respectively. The specific activity of trypsin enzyme in control susceptible colony was 20.806 +/- 0.452micromol/min/mg protein; but at F4 and F8 for malathion and methomyl treated larvae were 10.810 +/- 0.860 & 15.616+/-0.408 micromol/min/mg protein, respectively. Trypsin activity of F12 in treated larvae with B.t.H14 was 2.097 +/- 0.587 microiol/min/mg protein. Aminopeptidase specific activity for susceptible control larvae was 173.05 +/- 1.3111 micromol/min/mg protein. This activity decreased to 145.15 +/- 4.12, 152.497 +/- 6.775 & 102.04 +/- 3.58a micromol/min/mg protein after larval (F 12) treatment with methomyl, malathion and B.t H 14 respectively.

This paper is concerned mainly with the relationship between microfilarial periodicity and vector periodicity. The so-called ”non-periodic” Pacific form of Wuchereria bancrofti in fact shows a well-marked and relatively constant periodicity. The amplitude of this periodicity is low, which may account for the difficulty of detecting it in small clinical samples. The periodicity is well adapted to the biting cycle of Aëdes polynesiensis, though less so than that of the ”semi-periodic” Brugia malayi to forest Mansonioides. Microfilarial periodicity is discussed in the light of recent work on circadian rhythms in other animals and it is suggested that certain possibilities have been given insufficient weight. The use of the term ”migration” to describe microfilarial translocation within the host may have given rise to misunderstanding but it is in good agreement with current concepts of migration. Recent work suggests some behavioural heterogeneity in Culex fatigans which could render it a useful mosquito for elucidating certain important but little-understood problems. PMID:13933891

The toxicity of Pinus densiflora (red pine) hydrodistillate, its 19 constituents and 28 structurally related compounds against early third-instar larvae of Aedes albopictus (Ae. albopictus), Aedes aegypti (Ae. aegypti) and Culexpipiens palles (Cx. p. pallens) was examined using direct-contact bioassays. The efficacy of active compounds was further evaluated in semi-field bioassays using field-collected larval Cx. p. pallens. Results were compared with those of two synthetic larvicides, temephos and fenthion. In laboratory bioassays, Pinus densiflora hydrodistillate was found to have 24 h LC50 values of 20.33, 21.01 and 22.36 mg/L against larval Ae. albopictus, Ae. aegypti and Cx. p. pallens respectively. Among the identified compounds, thymol, δ-3-carene and (+)-limonene exhibited the highest toxicity against all three mosquito species. These active compounds were found to be nearly equally effective in field trials as well. In vitro bioassays were conducted to examine the acetylcholinesterase (AChE) inhibitory activity of 10 selected compounds. Results showed that there is a noticeable correlation between larvicidal activity and AChE inhibitory activity. In light of global efforts to find alternatives for currently used insecticides against disease vector mosquitoes, Pinus densiflora hydrodistillate and its constituents merit further research as potential mosquito larvicides.

Wolbachia is a maternally inherited endosymbiont inducing various effects in insects and other invertebrate hosts that facilitate the invasion of naive host populations. One of the effects is a form of sterility known as cytoplasmic incompatibility (CI) through which females are effectively sterilized when they mate with males harboring a different Wolbachia strain. The repeated mass release of cytoplasmically incompatible males can be a tool to suppress insect populations. Here, we attempt to infect an Aedes albopictus (Skuse) (Diptera: Culicidae) strain, artificially deprived of the natural Wolbachia infection, with a new Wolbachia strain from Culexpipiens (L.) (Diptera: Culicidae). Further experiments were designed to study the effects of the new infection on Ae. albopictus fitness and evaluate key parameters that affect infection dynamics, including CI level and maternal inheritance. Using embryonic microinjection, the new Wolbachia strain was successfully established in Ae. albopictus. Crosses demonstrated a pattern of bidirectional CI between naturally infected and transinfected individuals. Specifically, egg hatch was essentially absent (i.e., CI was very high) in all crosses between the transinfected males and females with a different infection status. Furthermore, naturally infected Ae. albopictus males were incompatible with the transinfected females. Maternal inheritance was close to 100%. Moreover, the new infection did not affect immature and adult survivorship, but it significantly reduced female fecundity and egg hatch rate. The results are discussed in relation to the potential use of the new Ae. albopictus-Wolbachia symbiotic association as a suitable system for the study and development of CI-based strategies for suppressing populations of this important pest and disease vector.

We report that two laboratory colonies of Culex quinquefasciatus and Culexpipiens mosquitoes were experimentally unable to transmit ZIKV either up to 21 days post an infectious blood meal or up to 14 days post intrathoracic inoculation. Infectious viral particles were detected in bodies, heads or saliva by a plaque forming unit assay on Vero cells. We therefore consider it unlikely that Culex mosquitoes are involved in the rapid spread of ZIKV.

We report that two laboratory colonies of Culex quinquefasciatus and Culexpipiens mosquitoes were experimentally unable to transmit ZIKV either up to 21 days post an infectious blood meal or up to 14 days post intrathoracic inoculation. Infectious viral particles were detected in bodies, heads or saliva by a plaque forming unit assay on Vero cells. We therefore consider it unlikely that Culex mosquitoes are involved in the rapid spread of ZIKV. PMID:27605159

Mosquitoes of the Culexpipiens group, including Culex quinquefasciatus, are important vectors of West Nile virus and other viruses around the world. Control of these mosquitoes in urban areas is often difficult because of the use of storm drains and other man-made structures as larval habitats. I...

The occurrence of multiple pathogens in mosquitoes and birds could affect the dynamics of disease transmission. We collected adult Culexpipiens and Cx. restuans (Cx. pipiens/restuans hereafter) from sites in Wisconsin and tested them for West Nile virus (WNV) and for avian malaria (Plasmodium). Gravid Cx. pipiens/restuans were tested for WNV using a commercial immunoassay, the RAMP?? WNV test, and positive results were verified by reverse transcriptasepolymerase chain reaction. There were 2 WNV-positive pools of Cx. pipiens/restuans in 2006 and 1 in 2007. Using a bias-corrected maximum likelihood estimation, the WNV infection rate for Cx. pipiens/restuans was 5.48/1,000 mosquitoes in 2006 and 1.08/1,000 mosquitoes in 2007. Gravid Cx. pipiens or Cx. restuans were tested individually for avian Plasmodium by a restriction enzymebased assay. Twelve mosquitoes were positive for avian Plasmodium (10.0), 2 were positive for Haemoproteus, and 3 were positive for Leucocytozoon. There were 4 mixed infections, with mosquitoes positive for >1 of the hemosporidian parasites. This work documents a high rate of hemosporidian infection in Culex spp. and illustrates the potential for co-infections with other arboviruses in bird-feeding mosquitoes and their avian hosts. In addition, hemosporidian infection rates may be a useful tool for investigating the ecological dynamics of Culex/avian interactions. ?? 2010 by The American Mosquito Control Association, Inc.

The abundance and seasonal distribution of Culex mosquitoes were monitored by measuring oviposition activity during the summers of 1995, 1996, and 1997. Five species of culicine mosquitoes laid egg rafts in ovitraps. Egg rafts of 4 Culex species-Cx. restuans (54.98%), Cx. pipiens (25.41%), Cx. salinarius (12.18%), and Cx. tarsalis (0.14%)-constituted more than 92% of the total egg rafts collected. Culiseta inornata (0.1%) was the only other species to lay viable egg rafts in the ovitraps. A small percentage (7.19%) of egg rafts did not hatch; thus identification was not possible. Compared with New Jersey light trap data in a nearby area, the abundance of Cx. tarsalis and Cs. inornata was markedly underestimated with ovitraps. These data may also reflect differences in the specific location of the 2 trap sites rather than attractiveness of ovitraps for Cx. tarsalis and Cs. inornata. In general, Cx. restuans oviposition activity began in late May, and it was the dominant Culex species through June. After July 1, the number of egg rafts laid by Cx. restuans decreased continually until the end of the summer. Culexpipiens and Cx. salinarius oviposition began in early June and increased gradually during the summer. Over the course of a summer, there were about twice as many Cx. pipiens as there were Cx. salinarius. A crossover in the number of Cx. pipiens and Cx. restuans egg rafts occurred in late July or early August, depending on the year. By late August each year, Cx. pipiens was the most abundant species. At this time, Cx. restuans and Cx. salinarius populations were similar, but each was about half of the Cx. pipiens population. Differences in the abundance of all 3 species could not be explained by changes in ambient temperature (both minimum and maximum) or relative humidity, either within or among years.

A major expenditure in vector biology laboratories is the rearing of mosquitoes. Most mosquito colonies require substantial effort to maintain, including frequent blood meals for optimal performance. Successful cryopreservation of mosquitoes continues to be elusive. Although using diapause as a s...

The continuous culture of mosquitoes is a costly endeavor for vector biology laboratories. In addition to the resources that must be committed to colony maintenance, biological costs including genetic drift and accidental colony loss can also occur. While alternatives do exist, their application t...

Culex species were monitored at three proximate sites with historically different West Nile virus (WNV) activities. The site with human WNV transmission (epidemic) had the lowest abundance of the putative bridge vectors, Culexpipiens and Cx. salinarius. The site with horse cases but not human cases (epizootic) had the highest percent composition of Cx. salinarius, whereas the site with WNV-positive birds only (enzootic) had the highest Cx. pipiens abundance and percent composition. A total of 29 WNV-positive Culex pools were collected at the enzootic site, 17 at the epidemic site, and 14 at the epizootic site. Published models of human risk using Cx. pipiens and Cx. salinarius as the primary bridge vectors did not explain WNV activity at our sites. Other variables, such as additional vector species, environmental components, and socioeconomic factors, need to be examined to explain the observed patterns of WNV epidemic activity.

Climate-based models simulating Culex mosquito population abundance in the Northeastern US were developed. Two West Nile vector species, Culexpipiens and Culex restuans, were included in model simulations. The model was optimized by a parameter-space search within biological bounds. Mosquito population dynamics were driven by major environmental factors including temperature, rainfall, evaporation rate and photoperiod. The results show a strong correlation between the timing of early population increases (as early warning of West Nile virus risk) and decreases in late summer. Simulated abundance was highly correlated with actual mosquito capture in New Jersey light traps and validated with field data. This climate-based model simulates the population dynamics of both the adult and immature mosquito life stage of Culex arbovirus vectors in the Northeastern US. It is expected to have direct and practical application for mosquito control and West Nile prevention programs.

Climate-based models simulating Culex mosquito population abundance in the Northeastern US were developed. Two West Nile vector species, Culexpipiens and Culex restuans, were included in model simulations. The model was optimized by a parameter-space search within biological bounds. Mosquito population dynamics were driven by major environmental factors including temperature, rainfall, evaporation rate and photoperiod. The results show a strong correlation between the timing of early population increases (as early warning of West Nile virus risk) and decreases in late summer. Simulated abundance was highly correlated with actual mosquito capture in New Jersey light traps and validated with field data. This climate-based model simulates the population dynamics of both the adult and immature mosquito life stage of Culex arbovirus vectors in the Northeastern US. It is expected to have direct and practical application for mosquito control and West Nile prevention programs.

La Crosse virus (LACV), a leading cause of arboviral pediatric encephalitis in the United States, is emerging in Appalachia. Here, we report field and laboratory evidence that suggest LACV may be using Culex mosquitoes as additional vectors in this region. This bunyavirus was detected by reverse-transcriptase polymerase chain reaction in two pools of Culex mosquitoes in southwestern Virginia and in six pools in West Virginia. To assess vector competence, we offered LACV blood meals to field-collected Culex restuans Theobald, Cx. pipiens L., and Aedes triseriatus (Say). Both Culex species were susceptible to infection. LACV-positive salivary expectorate, indicative of the ability to transmit, was detected in a small proportion of Cx. restuans (9%) and Cx. pipiens (4%) compared with Ae. triseriatus (40%). In a companion study of Cx. restuans only, we found that adults derived from nutritionally stressed larvae were significantly more likely to disseminate and transmit LACV. Our results indicate a potential role of Culex spp. in LACV dynamics that should be explored further in endemic areas. PMID:26175029

Wolbachia are α-proteobacteria that were first reported in Culexpipiens mosquitoes early in the twentieth century. Since then, the effect of Wolbachia on their host's reproduction has drawn attention and has been increasingly investigated. Given the extreme complexity of this interaction, new study cases are welcomed to enhance its understanding. The present work addressed the influence of Wolbachia on Cx. quinquefasciatus, the cosmopolitan member of the Cx. pipiens complex. Samples of a Cx. quinquefasciatus colony (wPip(+)) originated from individuals naturally infected by Wolbachiapipientis B strain, were cured with tetracycline, yielding a Wolbachia-free colony (wPip(-)). Both the presence of bacteria and the efficiency of bacterial elimination were checked by PCR of the wsp gene. Total reproductive unidirectional incompatibility occurred when wPip(-) females were crossed with wPip(+) males, whereas the other three types of reciprocal crosses were viable. Reproductive aspects were also comparatively evaluated between colonies. Concerning oviposition time during the first gonotrophic cycle, wPip(+) females developed and laid eggs earlier than did wPip(-) females. Reproductive fitness was higher among wPip(-) than wPip(+) females regarding the following parameters: fertility: egg rafts/fed females; fecundity: eggs/raft, and viability: larvae/eggs. Conversely, longevity of wPip(-) females was lower. Summarising, although the infected mosquitoes have the advantage of a higher longevity, they have lower reproductive fitness. Our results are partly distinct from all other reports on Aedes and Culex mosquitoes previously published.

Given their medical and veterinary relevance, the members of the Pipiens Assemblage are a worldwide target of ecological research. The distribution of Culexpipiens s.s. and Cx. quinquefasciatus converge in Buenos Aires, Argentina, where hybrids have been detected. Each member of the assemblage exhibits a distinct eco-physiological behaviour that can affect its efficiency in pathogen transmission. Our aim was to identify the environmental drivers for the spatio-temporal distribution of each member, focusing on latitudinal and urbanisation gradients. Immatures of mosquitoes were surveyed in artificial containers found within 11 public cemeteries, raised up to the adult stage and identified by their male genitalia. The distribution of each member was associated with the environment in a Generalized Linear Model. The variable accounting for most of the heterogeneity was latitude; Cx. quinquefasciatus was collected more frequently at northern cemeteries, whereas Cx. pipiens and hybrids were more likely at the southern extreme. The urbanisation gradient was also associated with the occurrence of Cx. quinquefasciatus and hybrids at the high and low end, respectively. Other relevant variables were cemetery total area, the proportion with graves and the presence of plastic flowers in the containers. The spatial distribution of the members of the Pipiens Assemblage within the sympatric region in South America is driven by environmental features. The information presented herein provides essential baseline data for surveillance programs and control activities.

Given their medical and veterinary relevance, the members of the Pipiens Assemblage are a worldwide target of ecological research. The distribution of Culexpipiens s.s. and Cx. quinquefasciatus converge in Buenos Aires, Argentina, where hybrids have been detected. Each member of the assemblage exhibits a distinct eco-physiological behaviour that can affect its efficiency in pathogen transmission. Our aim was to identify the environmental drivers for the spatio-temporal distribution of each member, focusing on latitudinal and urbanisation gradients. Immatures of mosquitoes were surveyed in artificial containers found within 11 public cemeteries, raised up to the adult stage and identified by their male genitalia. The distribution of each member was associated with the environment in a Generalized Linear Model. The variable accounting for most of the heterogeneity was latitude; Cx. quinquefasciatus was collected more frequently at northern cemeteries, whereas Cx. pipiens and hybrids were more likely at the southern extreme. The urbanisation gradient was also associated with the occurrence of Cx. quinquefasciatus and hybrids at the high and low end, respectively. Other relevant variables were cemetery total area, the proportion with graves and the presence of plastic flowers in the containers. The spatial distribution of the members of the Pipiens Assemblage within the sympatric region in South America is driven by environmental features. The information presented herein provides essential baseline data for surveillance programs and control activities. PMID:27783720

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex ( Culex ) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species ( Cx. acharistus , Cx. chidesteri , Cx. dolosus , Cx. lygrus and Cx. saltanensis ) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus . Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex ( Cux. ) species. PMID:24473810

Culexpipiens s.l. is one of the primary vectors of West Nile Virus in the USA and Continental Europe. The seasonal abundance and eco-behavioural characteristics of the typical form, Cx. pipienspipiens, make it a key putative vector in Britain. Surveillance of Culex larvae and adults is essential to detect any changes to spatial and seasonal activity or morphological traits that may increase the risk of disease transmission. Here we report the use of the modified Reiter gravid box trap, which is commonly used in the USA but scarcely used in the UK, to assess its suitability as a tool for British female Culex mosquito surveillance. Trapping was carried out at 110 sites in urban and rural gardens in Berkshire in May, July and September 2013. We tested if reproductively active adult female Culex are more abundant in urban than rural gardens and if wing characteristic traits and egg raft size are influenced by location and seasonal variations. Gravid traps were highly selective for Culex mosquitoes, on average catching significantly more per trap in urban gardens (32.4 ± 6.2) than rural gardens (19.3 ± 4.0) and more in July than in May or September. The majority of females were caught alive in a good condition. Wing lengths were measured as an indicator of size. Females flying in September were significantly smaller than females in May or July. Further non-significant differences in morphology and fecundity between urban and rural populations were found that should be explored further across the seasons.

Difficulties in correctly differentiating Culex restuans mosquitoes from Culexpipiens have left the spatiotemporal mechanisms underlying the epidemiology of West Nile virus (WNV) in the northeastern United States largely unresolved. We performed weekly surveys across a natural to urban gradient of sites in central New Jersey (USA) and used a rapid and cheap DNA extraction and a species-specific PCR assay to create single species pools for WNV testing. To assess seasonal trends we combined these results with WNV surveillance records generated from grouped Cx. restuans/Cx. pipiens pools tested in 2011-2012. Cx.restuans was found to be highly abundant within all sites and reached especially high abundance in urban wetland habitats greatly disturbed by human action. In contrast, the seasonal presence of Cx. pipiens was greatest in residential and urban habitats and its presence in natural areas was minimal throughout the season. WNV infection rates in both species were similar but Cx. restuans was consistently found infected first and more frequently, even as early as May, whereas WNV was first detected in Cx. pipiens in late July. WNV activity peaked during the month of August when WNV was commonly isolated from both species. The peak in WNV activity in August observed for both species was consistent with data from 2011 to 2012 when Cx. restuans and Cx. pipiens were grouped, although analyzing single species pools increased overall predicted infection levels. Our results support the preeminence of Cx. restuans as an enzootic vector of WNV and strongly suggest this species has become a "native invasive" exploiting human modified habitats and reaching very high abundance there. Importantly, high infection rates in disturbed wetland sites with high populations of Cx. restuans suggest this species may enable the introduction of WNV to urbanized environments where both Culex contribute to transmission potentiating disease risk.

Climatic changes forecasted in the coming years are likely to result in substantial alterations to the distributions and populations of vectors of arthropod-borne pathogens. Characterization of the effect of temperature shifts on the life history traits of specific vectors is needed to more accurately define how such changes could impact the epidemiological patterns of vector-borne disease. Here, we determined the effect of temperatures including 16, 20, 24, 28, and 32 degreeC on development time, immature survival, adult survival, mosquito size, blood feeding, and fecundity of both field and colonized populations of the Culex mosquitoes Culexpipiens L, Culex quinquefasciatus Say, and Culex restuans Theobald. Our results demonstrate that temperature significantly affects all of these traits, yet also that the extent of this effect is at times incongruent among temperatures, as well as being population and species-specific. Comparisons of colonized mosquitoes with field populations generally demonstrate decreased adult and immature survival, increased blood feeding and egg production, and significant variation in the effects of temperature, indicating that such colonies are not fully representative of natural populations. Results with field populations in general indicate that increases in temperature are likely to accelerate mosquito development, and that this effect is greater at temperatures below 24 degreeC, but also that temperature significantly increases mortality. Among field populations, Cx. restuans were most affected by temperature increases, with decreased longevity relative to other species and significant increases in adult and immature mortality measured with each incremental temperature increase. Despite the unique climates characteristic of the geographic ranges ofCx. quinquefasciatus and Cx. pipiens, evidence of significant species-specific adaptation to temperature ranges was not seen. Taken together, these results indicate that geographic region

A widely recommended strategy to minimize mosquito production in structural stormwater Best Management Practices (BMPs) is to ensure they hold captured water for no more than 72 h. However, this standard may be overly conservative for many mosquito species found in urban environments and may impede or prevent the capacity of BMPs to fulfill more stringent water quality standards in environmentally sensitive areas. Egg-to-pupa development of Culex tarsalis, Cx. pipiens, and Cx. quinquefasciatus were examined during July, August, and September 2006 in stormwater management basins and in water collected from these basins in 3 climatically distinct regions of California: the Lake Tahoe Basin, Sacramento Valley, and Los Angeles Basin. The observed minimum times to pupal development were 6 days for Cx. tarsalis and Cx. quinquefasciatus and 8 days for Cx. pipiens. Multiple linear regression models were used to estimate minimum predicted development times under optimal conditions for each region. The results suggest that water residence times of up to 96 h will not significantly increase the potential for Culex mosquito production in stormwater BMPs in the 3 regions included in this study.

Interspecific variations in the susceptibility of freshly and embryonated eggs of Aedes albopictus, Ae. aegypti, Ae. atropalpus and Culexpipiens were tested against three classes of insect growth regulators (IGRs) including ecdysone agonist (azadirachtin), chitin synthesis inhibitor (diflubenzuron) and juvenile hormone analog (pyriproxyfen) at 0.001, 0.01, 0.1 and 1.0ppm concentrations. Egg hatching inhibition was dose dependent, the highest being at 1.0ppm concentration for freshly laid eggs of Ae. albopictus (pyriproxyfen: 80.6%, azadirachtin: 42.9% and diflubenzuron: 35.8%). Aedes aegypti showed lower egg hatching inhibition when exposed to pyriproxyfen (47.3%), azadirachtin (15.7%) and diflubenzuron (25.5%). Freshly laid eggs of Cx. pipiens were most susceptible to diflubenzuron. Aedes atropalpus eggs were tolerant to all three classes of IGRs. Embryonated eggs of Ae. albopictus, Ae. aegypti, Ae. atropalpus and Cx. pipiens were resistant to pyriproxyfen, azadirachtin and diflubenzuron than freshly laid eggs. The median desiccation time (DT50) of Ae. atropalpus eggs was maximum (5.1h) as compared to Ae. aegypti (4.9h), Ae. albopictus (3.9h) or Cx. pipiens (1.7h) eggs. Insignificant relationship between the rates of desiccation and egg hatching inhibition suggests other factors than physical providing eggs the ability to tolerate exposures to various IGRs. Egg hatching inhibition was due to the alteration in embryonic development caused by IGRs. Changes in the egg shell morphology and abnormal egg hatching from the side of the egg wall instead of operculum, was observed at higher concentrations of diflubenzuron. Morphological and physiological variations in eggs may be the key factor to influence the ovicidal efficacy of IGRs. The present data provide a base line for the improvement of the ovicidal efficacy of the insecticide and its formulation.

The mosquito iridescent viruses (MIVs) are large icosahedral DNA viruses that replicate and assemble in the cytoplasm of the host. Paracrystalline arrangements of virions that accumulate in the cytoplasm produce an iridescent color that is symptomatic of acute infections. In August 2010, we found ...

One major obstacle in vector biology experimentation is the rearing of mosquitoes. Most mosquito colonies require substantial effort to maintain, including a blood meal at least once a month for optimal performance. While the induction of diapause can be used to reduce the amount of work required ...

The kinematics and hydrodynamics of free-swimming pupal and larval (final-instar) culicids were investigated using videography and a simple wake-visualisation technique (dyes). In both cases, swimming is based on a technique of high-amplitude, side-to-side (larva) or up-and-down (pupa) bending of the body. The pupa possesses a pair of plate-like abdominal paddles; the larval abdominal paddle consists of a fan of closely spaced bristles which, at the Reynolds numbers involved, behaves like a continuous surface. Wake visualisation showed that each half-stroke of the swimming cycle produces a discrete ring vortex that is convected away from the body. Consecutive vortices are produced first to one side then to the other of the mean swimming path, the convection axis being inclined at approximately 25 degrees away from dead aft. Pupal and larval culicids therefore resemble fish in using the momentum injected into the water to generate thrust. Preliminary calculations for the pupa suggest that each vortex contains sufficient momentum to account for that added to the body with each half-stroke. The possibility is discussed that the side-to-side flexural technique may allow an interaction between body and tail flows in the production of vorticity.

Ndumu virus (NDUV) is a member of the Family: Togaviridae and Genus: Alphavirus. In Kenya the virus has been isolated from a range of mosquito species but has not been associated with human or animal morbidity. Little is know about the transmission dynamics or vertebrate reservoirs of this virus. We...

Reassortant viruses replicat- < Valley fever (RVF) virus . The origin of the S and M ed efficiently in mosquitoes and were readily transmis- RNA segments...epidemiology and evolution of RVF parental viruses by interrupted feeding on two infected virus . Introduction between spontaneous mutants arising in an infected...of RVF virus , in mosquitoes that fed on a vertebrate host 1990). As is well documented with other viruses infected with two different strains of RVF

One specimen of a mosquito new to Florida, Culex declarator, was first found in 1998 in Indian River County. A 2nd specimen was collected in 2002. Beginning in September 2003, Cx. declarator adults were regularly encountered in routine mosquito surveillance sampling, with more than 300 specimens appearing in 45 collections. Prior to our find, the U.S. distrubution was thought to be restricted to south Texas. The full extent of this species' distribution in Florida has yet to be determined.

West Nile virus is a mosquito borne flavivirus endemic over a large geographic area including Africa, Asia, and the Middle East. Although the virus generally causes a mild, self-limiting febrile illness in humans, it has sporadically caused central nervous system infections during epidemics. An isolate of West Nile virus was obtained from a pool of four male Culex univittatus complex mosquitoes while we were conducting an investigation of Rift Valley fever along the Kenya-Uganda border in February-March 1998. This represents the first field isolation of West Nile virus from male mosquitoes and strongly suggests that vertical transmission of the virus occurs in the primary maintenance mosquito vector in Kenya. A phylogenetic analysis of the complete amino acid sequence of the viral envelope glycoprotein demonstrated a sister relationship with a Culexpipiens mosquito isolate from Romania made in 1996. This unexpected finding probably reflects the role of migratory birds in disseminating West Nile virus between Africa and Europe.

Culex tarsalis Coquillett, Cx. quinquefasciatus Say, and Cx. pipiens L. were collected during the warm winter of 2009 using dry ice-baited and gravid traps and walk-in red boxes positioned in desert, urban, and agricultural habitats in Riverside, Los Angeles, Kern, and Yolo Counties. Temperatures exceeded the preceding 50 yr averages in all locations for most of January, whereas rainfall was absent or below average. Abundance of Culex species in traps during January ranged from 83 to 671% of the prior 5 yr average in all locations. Few females collected resting were in diapause during January based on follicular measurements. Evidence for early season gonotrophic activity included the detection of freshly bloodfed, gravid, and parous females in resting collections, gravid oviposition site-seeking females in gravid female traps, and nulliparous and parous host-seeking females at dry ice-baited traps. Female Culex seemed to employ multiple overwintering strategies in California, including larval and adult quiescence, adult female diapause, and an intermediate situation with adult females collected with enlarged follicles, but without evident vitellogenesis. West Nile, St. Louis, or western equine encephalitis viruses were not detected in 198 pools of adults or 56 pools of adults reared from field-collected immatures collected during January and February 2009. Our preliminary data may provide insight into how climate change may extend the mosquito season in California.

Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect of its co-infection on vector competence for Dirofilaria immitis (Spirurida: Onchocercidae) in the wild has not been investigated. This study aimed to screen vectors of D. immitis for wPip, to characterize these, and to investigate a possible association between the occurrence of W. pipientis and that of the nematode. The presence of W. pipientis was assessed in the five mosquito potential vectors of D. immitis in Portugal. Polymerase chain reaction (PCR) products were sequenced, and wPip haplotypes were determined by PCR-restricted fragment length polymorphism (RFLP). Results showed that wPip was detected in 61.5% of Culexpipiens (Diptera: Culicidae) pools and 6.3% of Culex theileri pools. wPip 16s rRNA sequences found in Cx. theileri exactly match those from Cx. pipiens, confirming a mosquito origin, rather than a nematode origin, as some specimens were infected with D. immitis. Only wPip haplotype I was found. No association was found between the presence of wPip and D. immitis in mosquitoes and hence a role for this endosymbiont in influencing vectorial competence is yet to be identified. This study contributes to understanding of wPip distribution in mosquito populations and, to the best of the authors' knowledge, is the first report of natural infections by wPip in Cx. theileri.

Study of statistically significant rhythms of mitotic division in dorsal epidermis of frogs, Rana pipiens, exposed to a 12:12 light:dark environment for 14 days. The results include the findings that (1) male animals have a primary period of 22 hr in summer and 18 hr in winter, (2) female animals have an 18 hr period, and (3) parapinealectomy and blinding abolish the rhythm.

The main tendency for the control of West Nile virus vectors, without the presence of disease, is to perform integrated programs minimizing chemicals by using environmentally friendly substances which act as oviposition attractants such as oviposition pheromones and infusions. This is the first time that an aged infusion is combined with aged pheromone (microencapsulated). Initially, three common plants in Greece were evaluated as a potential oviposition medium: Oxalis pes-carpae, Jasminum polyanthum, and Avena barbata. All revealed an excellent oviposition attractancy which was more than 80%. O. pes-carpae was used for further investigation and attractancy over time was also studied. Finally, the combination of the synthetic pheromone (6-acetoxy-5-hexadecanolide) with the O. pes-carpae infusion revealed a synergistic effect only for the first day. This project was a first detection for the potential use of microencapsulated synthetic pheromone with infusion and results are discussed.

Marker dyes are often mixed with liquid insecticide formulations prior to field applications to accurately determine the characteristics and penetration of droplets into targeted habitats. We have been using FD&C Red 40 Granular DM food dye at the rate of 20 g/liter in liquid solutions of Bacillus thuringiensis israelensis (Bti) for area-wide larvicide applications against the Asian tiger mosquito, Aedes albopictus. The Bti and dye mix ratio has been recommended by pesticide manufacturers for testing under operational conditions, but no data exist on the effects of the dye itself on mosquito larvae. We tested the effects of the FD&C Red 40 food dye in laboratory bioassays against different strains of Ae. albopictus (New Jersey and Maryland) and Culexpipienspipiens (Utah) at rates of 0.039 to 80.0 g/liter. We also conducted field application trials to measure dye concentrations up to 100 m downwind when mixed and applied according to manufacturer instructions. In laboratory bioassays, we found that mean survival in cups with dye were significantly different from the controls beginning at 10.0 g/liter for New Jersey Ae. albopictus and at 20.0 g/liter for Maryland Ae. albopictus and Utah Cx. p. pipiens. In field application trials, we recorded a maximum volume density of 1,152.8 nl/cm(2) and calculated the maximum concentration of dye at 9.09 × 10(-3) g/liter. Our results showed that although we detected greater effects of dye on Ae. albopictus in New Jersey experiments than Ae. albopictus in Maryland and Cx. p. pipiens from Utah, concentrations of the dye during operational applications were at least 1,100 times below concentrations that exhibited toxic effects for either species in the laboratory, suggesting that the dye will not interfere with accuracy of field bioassays. Our results conclusively demonstrate that the addition of the FD&C Red 40 marker dye does not alter the efficacy of the pesticide formulation by skewing results, but rather provides a valuable

Male and female adults of Culex (Culex) articularis Philippi are redescribed, and the 4th-instar larva and pupa are described and illustrated for the first time. Culex articularis is compared with other species of the subgenus Culex. Illustrations of diagnostic characters of the female, male genitalia, 4th-instar larva, and pupa are also provided.

The pupa of Culex declarator was partially described and illustrated by Lane. This species was recently discovered in Florida for the first time and a series of individual rearings have resulted in a number of pupae and an opportunity to redescribe it fully, with a complete illustration. Amendment to the key to the Culex pupae of the eastern United States is provided.

Formal publication of the Culex genome sequence has closed the human disease vector triangle by meeting the Anopheles gambiae and Aedes aegypti genome sequences. Compared to these other mosquitoes, Culex quinquefasciatus possesses many specific hallmark characteristics, and may thus provide different angles for research which ultimately leads to a practical solution for controlling the ever increasing burden of insect-vector-borne diseases around the globe. We argue the special importance of the cosmopolitan species- Culex genome sequence by invoking many interesting questions and the possible of potential of the Culex genome to answer those.

An accurate understanding and prediction of mosquito population dynamics are needed to identify areas where there is a high risk of mosquito-borne disease spread and persistence. Simulation tools are relevant for supporting decision-makers in the surveillance of vector populations, as models of vector population dynamics provide predictions of the greatest risk periods for vector abundance, which can be particularly helpful in areas with a highly variable environment. We present a generic weather-driven model of mosquito population dynamics, which was applied to one species of each of the genera Anopheles, Culex, and Aedes, located in the same area and thus affected by similar weather conditions. The predicted population dynamics of Anopheles hyrcanus, Culexpipiens, and Aedes caspius were not similar. An. hyrcanus was abundant in late summer. Cx. pipiens was less abundant but throughout the summer. The abundance of both species showed a single large peak with few variations between years. The population dynamics of Ae. caspius showed large intra- and inter-annual variations due to pulsed egg hatching. Predictions of the model were compared to longitudinal data on host-seeking adult females. Data were previously obtained using CDC-light traps baited with carbon dioxide dry ice in 2005 at two sites (Marais du Viguerat and Tour Carbonnière) in a favourable temperate wetland of southern France (Camargue). The observed and predicted periods of maximal abundance for An. hyrcanus and Cx. pipiens tallied very well. Pearson's coefficients for these two species were over 75% for both species. The model also reproduced the major trends in the intra-annual fluctuations of Ae. caspius population dynamics, with peaks occurring in early summer and following the autumn rainfall events. Few individuals of this species were trapped so the comparison of predicted and observed dynamics was not relevant. A global sensitivity analysis of the species-specific models enabled us to

Rana pipiens larvae were exposed for 48 h in a flow-through system to clean water or five concentrations of the phototoxic polycyclic aromatic hydrocarbon (PAH) fluoranthene. Following this uptake period, the larvae were divided into four groups: one for immediate tissue residue analysis, a second for residue analysis following 48 h of depuration in clean water, and two for a 48-h exposure in clean water to ultraviolet (UV) light at two different levels. At the highest treatment, mean intensity was 8.12 {+-} 0.19 {times} 10{sup 2} {micro}W/cm{sup 2}, whereas at a lower treatment the UVA intensity was 4.45 {+-} 0.05 {times} 10{sup 2} {micro}W/cm{sup 2}. Larval frogs bioaccumulated fluoranthene in direct proportion to the water exposure concentrations, with initial whole-body PAH concentrations of 1.48, 3.53, 4.85, 11.3, and 18.7 {micro}g/g at the five treatment levels. No mortality of the animals occurred during the 48-h uptake phase. When the frogs were placed in clean water, the fluoranthene was rapidly depurated, with up to 80% lost in 48 h. Exposure to UV light following fluoranthene exposure significantly enhanced toxicity of the PAH. Median time to death decreased as the product of UVA light intensity and fluoranthene body residue increased. For larval R. Pipiens, sufficient tissue residues of fluoranthene were bioaccumulated within 48 h, at water exposure concentrations in the range of 2 to 10 {micro}g/L, to be lethal when combined with a UVA exposure simulating a fraction of summertime, midday sunlight in northern latitudes.

The pupa of Culex restuans is redescribed in detail with a chaetotaxal table and a full illustration. The chaetotaxy of the pupa of Cx. restuans is compared with that of Cx. nigripalpus, the primary vector of St. Louis encephalitis and West Nile virus in Florida.

Incompatible insect techniques are potential methods for controlling Culex quinquefasciatus and avian disease transmission in Hawai‘i without the use of pesticides or genetically modified organisms. The approach is based on naturally occurring sperm-egg incompatibilities within the Culexpipiens complex that are controlled by different strains of the bacterial endosymbiont Wolbachia pipientis (wPip). Incompatibilities can be unidirectional (crosses between males infected with strain A and females infected with strain B are fertile, while reciprocal crosses are not) or bidirectional (reciprocal crosses between sexes with different wPip strains are infertile). The technique depends on release of sufficient numbers of male mosquitoes infected with an incompatible wPip strain to suppress mosquito populations and reduce transmission of introduced avian malaria (Plasmodium relictum) and Avipoxvirus in native forest bird habitats. Both diseases are difficult to manage using more traditional methods based on removal and treatment of larval habitats and coordination of multiple approaches may be needed to control this vector. We characterized the diversity of Wolbachia strains in C. quinquefasciatus from Hawai‘i, Kaua‘i, Midway Atoll, and American Samoa with a variety of genetic markers to identify compatibility groups and their distribution within and between islands. We confirmed the presence of wPip with multilocus sequence typing, tested for local genetic variability using 16 WO prophage genes, and identified similarities to strains from other parts of the world with a transposable element (tr1). We also tested for genetic differences in ankyrin motifs (ank2 and pk1) which have been used to classify wPip strains into five worldwide groups (wPip1–wPip5) that vary in compatibility with each other based on experimental crosses. We found a mixture of both widely distributed and site specific genotypes based on presence or absence of WO prophage and transposable

Background Models of the effects of environmental factors on West Nile virus disease risk have yielded conflicting outcomes. The role of precipitation has been especially difficult to discern from existing studies, due in part to habitat and behavior characteristics of specific vector species and because of differences in the temporal and spatial scales of the published studies. We used spatial and statistical modeling techniques to analyze and forecast fine scale spatial (2000 m grid) and temporal (weekly) patterns of West Nile virus mosquito infection relative to changing weather conditions in the urban landscape of the greater Chicago, Illinois, region for the years from 2004 to 2008. Results Increased air temperature was the strongest temporal predictor of increased infection in Culexpipiens and Culex restuans mosquitoes, with cumulative high temperature differences being a key factor distinguishing years with higher mosquito infection and higher human illness rates from those with lower rates. Drier conditions in the spring followed by wetter conditions just prior to an increase in infection were factors in some but not all years. Overall, 80% of the weekly variation in mosquito infection was explained by prior weather conditions. Spatially, lower precipitation was the most important variable predicting stronger mosquito infection; precipitation and temperature alone could explain the pattern of spatial variability better than could other environmental variables (79% explained in the best model). Variables related to impervious surfaces and elevation differences were of modest importance in the spatial model. Conclusion Finely grained temporal and spatial patterns of precipitation and air temperature have a consistent and significant impact on the timing and location of increased mosquito infection in the northeastern Illinois study area. The use of local weather data at multiple monitoring locations and the integration of mosquito infection data from

male genitalia of these species are described and distinguished. Bram (1967) synonymized Culex (Cu7exJ interfor Dyar, 1928, with Cx. (Cux. J bidens...Dyar, 1922, on the basis of similarity in the male genitalia. The lectotype was the only male of interfor available to Bram, and he apparently...properly assess the status of interfor . In 1983 we notice !! that males of bidens collected in Santa Fe Province, Argentina could be separated into two

North Florida is a transition zone between widespread Culex nigripalpus populations to the south and focal Culex salinarius populations to the north. Culex nigripalpus is a vector of St. Louis encephalitis (SLE) and eastern equine encephalitis (EEE) viruses in south Florida, while Cx. salinarius is a suspected New World vector of West Nile (WN) virus. Abundant vector populations are often a prerequisite for epidemic and epizootic transmission of arboviruses. Extensive SLE transmission has never been reported from north Florida, but sporadic WN transmission was reported there during the summer of 2001. The disparate flavivirus transmission patterns observed in north and south Florida may be due, in part, to the local geographical and seasonal distribution of Culex vectors. Here we report that from May 1991 to April 1994, Cx. salinarius was most commonly observed during the winter and spring in northeast Florida (Duval County), whereas Cx. nigripalpus was most abundant during the summer and autumn. An unusually mild spring in 1991 allowed Cx. nigripalpus to reproduce early in the year, resulting in a summer population that emerged more than 8 wks earlier than in 1992 and 1993. The 1991 Cx. nigripalpus population persisted through October, when SLE transmission was detected by sentinel chickens. Transmission of SLE was not detected in Duval County during 1992 or 1993. These data indicate that mild winter and spring conditions in north Florida may favor increased abundance and survival of Cx. nigripalpus in a region where this species is normally not abundant. A seasonal shift in population structure may increase the transmission risk of arboviruses for which Cx. nigripalpus is a competent vector, including SLE, WN, and EEE.

Culex (Melanoconion) panocossa is a suspected vector of Venezuelan equine encephalitis virus in Central America. Prior to this report, Cx. panocossa was known from Central America (Belize, Costa Rica, El Salvador, Guatemala, southern Mexico, Panama), northern South America (Colombia, Venezuela) and the Greater Antilles (Cuba and Jamaica). Larvae (n=5) and adults (n=4286) of Cx. panocossa were collected at two locations near Homestead, FL, which indicates substantial established populations of this probable vector species in the continental US. Since larvae of Cx. panocossa are associated with Pistia spp. (water lettuce), the distribution of this mosquito is likely to expand in Florida, where water lettuce is a major invasive plant in freshwater ecosystems. The putative establishment of Cx. panocossa in Florida is of significant concern from a public health perspective, as its proliferation in developed areas could link historically sylvatic transmission foci of Everglades virus with populated centers such as the greater Miami Metropolitan area.

A Hermes-based transposable element transformation system incorporating an enhanced green fluorescent protein (EGFP) marker was used to produce two transgenic lines of Culex quinquefasciatus (Say). The transformation frequency was approximately 12% and transformation of Culex was shown to be dependent on the presence of Hermes transposase. Injected Culex embryos were treated with four different heat shock regimes, two of which produced transformed individuals. These individuals were mated with wild-type mosquitoes and produced offspring which expressed the dominant EGFP gene in Mendelian ratios predicted for the stable integration of a gene at a single locus. The two transformed lines displayed distinct patterns of phenotypic expression, the expression of which has remained stable after fifteen generations. In these transgenic lines both the Hermes element and flanking plasmid DNA integrated into the Culex genome, as has been previously seen in Hermes-mediated transgenic strains of Aedes aegypti (L.). The high frequency of Culex transformation together with the dependence on the presence of Hermes transposase suggests that, as for Ae. aegypti, this mode of transposition into the germ-line genome occurs by an alternate mechanisms to the cut and paste type of transposition seen for this element in other insect species and in the somatic nuclei of mosquitoes. This is the first report of the genetic transformation of a species in the genus Culex and demonstrates that this medically important mosquito species can now, along with several other Culicine and Anopheline mosquito species, be genetically manipulated.

The Horn of Africa represents a region formerly known to be highly susceptible to mosquito-borne infectious diseases. In order to investigate whether autochthonous WNV transmission occurs in the Djibouti City area, in how far, and which of, the endemic Culex mosquito species are involved in WNV circulation activity,and whether sentinel site-enhanced near-real time surveillance (SSE-NRTS) may increase WNV detection sensitivity, mosquito vector monitoring was conducted from January 2010 to June 2012. Six monitoring locations, including two identified sentinel sites, considered most probable for potential anthroponotic and zoonotic virus circulation activity, have been continuously employed. Among the 20431 mosquitoes collected, 19069 (93.4%) were Cx. quinquefasciatus, and 1345 (6.6%) Cx. pipiens ssp. torridus. WNV lineage 2 circulation activity was detected between December 20th, 2010 and January 7th, 2011. Overall, 19 WNV RNA-positive mosquito pools were detected. Generally, urban environment-specific WNV-RNA circulation took place in Cx. pipiens ssp. torridus, whereas periurban and rural area-linked circulation was detected only in Cx. quinquefasciatus. Serological investigation data from 10 volunteers employed at the dislocated zoonotic WNV transmission sentinel site suggest that six persons (60%) had an acute, or recent, WNV infection. Results show that WNV should be considered endemic for Djibouti and sentinel site-enhanced near-real time surveillance is an elegant and highly effective epidemiological tool. In Djibouti, the endemicity level, public health impact and transmission modes of vector-borne diseases in concordance with locally optimized monitoring and control regimen deserve further investigation.

Bioactive compounds contained in the seed kernel and other parts of the neem tree (Azadirachta indica A. Juss) have been found to show insecticidal activities and other effects in many species of insects. These activities include antifeedancy, growth regulation, fecundity suppression, male sterility, oviposition repellency, changes in biological fitness such as loss of flying ability, immunodepression, enzyme inhibition, splitting of biological rhythms, and so forth. We investigated the ovicidal effects of various formulations of azadrirachtin (AZ) against the mosquitoes Culex tarsalis Coquillett and Culex quinquefasciatus Say. The formulations tested were wettable powder Azad WP10, emulsifiable concentrate Azad EC4.5, and technically pure AZ. The ovicidal activity of the test neem products was influenced by concentration of AZ, age of the egg rafts, and age of the neem preparations. Other factors such as formulation and mosquito species were also involved in the degree of ovicidal activity. When the egg rafts were deposited directly in fresh neem suspension and left there for 4 h before transfer to untreated water, 1 ppm of AZ produced almost 100% mortality in eggs. When egg rafts aged for 0, 4, 8, 12, and 24 h were exposed to 10 ppm neem suspensions for 36 h, the ovicidal activity was only attained in the egg rafts deposited directly (0 h old) in the neem suspension, not in those with ages of 4-24 h. On aging, depending on the formulations and mosquito species, the neem suspensions at 1 ppm completely lost ovicidal activity within 7-20 days. The egg rafts of Cx. quinquefasciatus were more susceptible to the test neem products than those of Cx. tarsalis. The formulated neem products were more persistent and effective than the technical AZ. The wettable powder (WP) formulation was slightly more persistent and effective than the emulsifiable concentrate (EC). The ovicidal activity of the neem products against mosquitoes from the current research clearly demonstrated

Wastewater effluents are complex mixtures containing a variety of anthropogenic compounds, many of which are known endocrine disruptors. In order to characterize the development and behavorial effects of such a complex mixture, northern leopard frogs, Rana pipiens, were e...

Ionic currents in immature, ovulated Rana pipiens oocytes (metaphase I) were studied using the voltage-clamp technique. At this stage of maturity the oocyte can produce action potentials in response to depolarizing current or as an "off response" to hyperpolarizing current. Reducing external Na+ to 1/10 normal (choline substituted) eliminated the action potentials and both the negative-slope region and zero-crossing of the I-V relation. Reducing external Cl- to 1/10 or 1/100 normal (methanesulfonate substituted) lengthened the action potential. The outward current was reduced and a net inward current was revealed. By changing external Na+, Cl-, and K+ concentrations and using blocking agents (SITS, TEA), three voltage- and time-dependent currents were identified, INa, IK and ICl. The Na+ current activated at about 0 mV and reversed at very positive values which decreased during maturation. Inward Na+ current produced the upstroke of the action potential. During each voltage-clamp step the Na+ current activated slowly (seconds) and did not inactivate within many minutes. The Na+ current was not blocked by TTX at micromolar concentrations. The K+ current was present only in the youngest oocytes. Because IK was superimposed on a large leakage current, it appeared to reverse at the resting potential. When leakage currents were subtracted, the reversal potential for IK was more negative than -110 mV in Ringer's solution. IK was outwardly rectifying and strongly activated above -50 mV. The outward K+ current produced an after hyperpolarization at the end of each action potential. IK was blocked completely and reversibly by 20 mM external TEA. The Cl- current activated at about +10 mV and was outwardly rectifying. ICl was blocked completely and reversibly by 400 microM SITS added to the bathing medium. This current helped repolarize the membrane following an action potential in the youngest oocytes and was the only repolarizing current in more mature oocytes that had lost

Before the development of magnetic resonance imaging (MRI), evoked potentials (EPs)-visual evoked potentials, somatosensory evoked potentials, and brain stem auditory evoked responses-were commonly used to determine a second site of disease in patients being evaluated for possible multiple sclerosis (MS). The identification of an area of the central nervous system showing abnormal conduction was used to supplement the abnormal signs identified on the physical examination-thus identifying the "multiple" in MS. This article is a brief overview of additional ways in which central nervous system (CNS) physiology-as measured by EPs-can still contribute value in the management of MS in the era of MRIs.

Culex quinquefasciatus mosquitoes have been successfully genetically modified only once, despite the efforts of several laboratories to transform and establish a stable strain. We have developed a transient gene expression method, in Culex, that delivers plasmid DNA directly to the mosquito haemolymph and additional tissues. We were able to express DsRed2 fluorescent protein in adult Cx. quinquefasciatus mosquitoes by injecting plasmids directly into their thorax. The expression of DsRed2 in adult Cx. quinquefasciatus mosquitoes is an important stepping stone to genetic transformation and the potential use of new control strategies and genetic interactions.

Music is strongly intertwined with memories-for example, hearing a song from the past can transport you back in time, triggering the sights, sounds, and feelings of a specific event. This association between music and vivid autobiographical memory is intuitively apparent, but the idea that music is intimately tied with memories, seemingly more so than other potent memory cues (e.g., familiar faces), has not been empirically tested. Here, we compared memories evoked by music to those evoked by famous faces, predicting that music-evoked autobiographical memories (MEAMs) would be more vivid. Participants listened to 30 songs, viewed 30 faces, and reported on memories that were evoked. Memories were transcribed and coded for vividness as in Levine, B., Svoboda, E., Hay, J. F., Winocur, G., & Moscovitch, M. [2002. Aging and autobiographical memory: Dissociating episodic from semantic retrieval. Psychology and Aging, 17, 677-689]. In support of our hypothesis, MEAMs were more vivid than autobiographical memories evoked by faces. MEAMs contained a greater proportion of internal details and a greater number of perceptual details, while face-evoked memories contained a greater number of external details. Additionally, we identified sex differences in memory vividness: for both stimulus categories, women retrieved more vivid memories than men. The results show that music not only effectively evokes autobiographical memories, but that these memories are more vivid than those evoked by famous faces.

The purpose of the present study was to assess the effect of crude extracts of marine actinobacteria on larvicidal, repellent, and ovicidal activities against Culex tritaeniorhynchus and Culex gelidus (Diptera: Culicidae). The early fourth instar larvae of C. tritaeniorhynchus and C. gelidus, reared in the laboratory, were used for larvicidal, ovicidal, and repellent assay with crude extracts of actinobacteria. Saccharomonospora spp. (LK-1), Streptomyces roseiscleroticus (LK-2), and Streptomyces gedanensis (LK-3) were identified as potential biocide producers. Based on the antimicrobial activity, three strains were chosen for larvicidal activity. The marine actinobacterial extracts showed moderate to high larvicidal effects after 24 h of exposure at 1,000 ppm and the highest larval mortality was found in extract of LK-3 (LC(50) = 108.08 ppm and LC(90) = 609.15 ppm) against the larvae of C. gelidus and (LC(50) = 146.24 ppm and LC(90) = 762.69 ppm) against the larvae of C. tritaeniorhynchus. Complete protections for 240 min were found in crude extract of LK-2 and LK-3 at 1,000 ppm, against mosquito bites of C. tritaeniorhynchus and C. gelidus, respectively. After 24-h treatment, mean percent hatchability of the ovicidal activity was observed. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Crude extracts of LK-1 and LK-3 showed no hatchability at 1,000 ppm against C. tritaeniorhynchus and C. gelidus, respectively. This is an ideal ecofriendly approach for the control of Japanese encephalitis vectors, C. tritaeniorhynchus and C. gelidus.

The most informative neurophysiological techniques available in the neurosurgical intensive care unit are electroencephalograph and somatosensory evoked potentials. Such tools, which give an evaluation of cerebral function in comatose patients, support clinical evaluation and are complementary to neuroimaging. They serve both diagnostic/prognostic and monitoring purposes. While for the former, discontinuous monitoring is sufficient, for the latter, to obtain increased clinical impact, continuous monitoring is necessary. To perform and interpret these examinations in the neurosurgical intensive care unit, both the technician and the neurophysiologist need specific training in the intensive care field. There is sufficient evidence to show that somatosensory evoked potentials are the best single indicator of early prognosis in traumatic and hypoxic-ischaemic coma compared to the Glasgow Coma Score, computed tomography scan and electroencephalograph. Indeed, somatosensory evoked potentials should always be combined with clinical examination to determine the prognosis of coma. Despite widespread use of somatosensory evoked potentials and their prognostic utility in acute brain injury, few studies exist on continuous somatosensory evoked potential monitoring in the intensive care unit. We carried out a pilot study of continuous electroencephalograph-somatosensory evoked potential monitoring in the neurosurgical intensive care unit (traumatic brain injury and intracranial haemorrhage, Glasgow Coma Score <9, intracranial pressure monitoring). All patients stable from a clinical and computed tomography scan point of view showed no significant somatosensory evoked potential modifications, while in the case of clinical deterioration (23%), somatosensory evoked potentials always showed significant modifications. While somatosensory evoked potentials correlated with short-term outcome, intracranial pressure showed a poor correlation. We believe neurophysiological monitoring is

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

In tests in Rangoon, Burma, of the larvicidal activity of various organophosphorus compounds against C. p. fatigans, it was found that the activity usually lasted much longer in septic tanks and pit latrines than in open drains. Dursban, Abate, fenthion and several other emulsifiable concentrates caused high larval mortality at a concentration of 0.05 ppm but 0.5 ppm was normally required to obtain a minimum of 1-2 weeks of complete larval control. For the desired residual activity, dosages about 40-400 times the laboratory LC95 values were normally required, depending on the compound used. The most effective formulation was Dursban emulsifiable concentrate, which, at a concentration of 0.5 ppm, was effective for 3, 7 and 12 weeks in concrete drains, pit latrines and septic tanks, respectively. Petroleum oils applied at rates of 25-40 US gal/ac (approx. 237 l/ha—380 l/ha) and a pyrethrum derivative at a concentration of 1.0 ppm were toxic to larvae but not highly residual. Some emulsifiable-concentrate/oil mixtures appeared to be outstandingly effective, although inconsistent results also occurred. Granular formulations were normally less effective than the emulsifiable concentrates. PMID:4253561

Zika virus is a newly emergent mosquito-borne flavivirus that has caused recent large outbreaks in the new world, leading to dramatic increases in serious disease pathology including Guillain-Barre syndrome, newborn microcephaly, and infant brain damage. Although Aedes mosquitoes are thought to be the primary mosquito species driving infection, the virus has been isolated from dozens of mosquito species, including Culex and Anopheles species, and we lack a thorough understanding of which mosquito species to target for vector control. We exposed Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes to blood meals supplemented with two Zika virus strains. Mosquito bodies, legs, and saliva were collected five, seven, and 14 days post blood meal and tested for infectious virus by plaque assay. Regardless of titer, virus strain, or timepoint, Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes were refractory to Zika virus infection. We conclude that Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes likely do not contribute significantly to Zika virus transmission to humans. However, future studies should continue to explore the potential for other novel potential vectors to transmit the virus.

The nocturnal mosquito, Culex quinquefasciatus,as a vector of West Nile virus is the target of many surveillance and control efforts. Surveillance of this species primarily consists of light traps baited with a variety of chemical lures. While much research has focused on optimization of the olfa...

Zika virus is a newly emergent mosquito-borne flavivirus that has caused recent large outbreaks in the new world, leading to dramatic increases in serious disease pathology including Guillain-Barre syndrome, newborn microcephaly, and infant brain damage. Although Aedes mosquitoes are thought to be the primary mosquito species driving infection, the virus has been isolated from dozens of mosquito species, including Culex and Anopheles species, and we lack a thorough understanding of which mosquito species to target for vector control. We exposed Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes to blood meals supplemented with two Zika virus strains. Mosquito bodies, legs, and saliva were collected five, seven, and 14 days post blood meal and tested for infectious virus by plaque assay. Regardless of titer, virus strain, or timepoint, Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes were refractory to Zika virus infection. We conclude that Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes likely do not contribute significantly to Zika virus transmission to humans. However, future studies should continue to explore the potential for other novel potential vectors to transmit the virus. PMID:28316896

In contrast to most local amphibian populations, northeastern populations of the Northern Leopard Frog (Lithobates pipiens) have displayed uncharacteristically high levels of genetic diversity that have been attributed to large, stable populations. However, this widely distributed species also occurs in areas known for great climatic fluctuations that should be reflected in corresponding fluctuations in population sizes and reduced genetic diversity. To test our hypothesis that Northern Leopard Frog genetic diversity would be reduced in areas subjected to significant climate variability, we examined the genetic diversity of L. pipiens collected from 12 sites within the Prairie Pothole Region of North Dakota. Despite the region's fluctuating climate that includes periods of recurring drought and deluge, we found unexpectedly high levels of genetic diversity approaching that of northeastern populations. Further, genetic structure at a landscape scale was strikingly homogeneous; genetic differentiation estimates (Dest) averaged 0.10 (SD = 0.036) across the six microsatellite loci we studied, and two Bayesian assignment tests (STRUCTURE and BAPS) failed to reveal the development of significant population structure across the 68 km breadth of our study area. These results suggest that L. pipiens in the Prairie Pothole Region consists of a large, panmictic population capable of maintaining high genetic diversity in the face of marked climate variability.

Morphological differentiation of mosquitoes in the subgenera Culex (Culex) and Culex (Phenacomyia) in Guatemala is difficult, with reliable identification ensured only through examination of larval skins from individually reared specimens and associated male genitalia. We developed a multiplexed polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay to identify common Cx. (Cux.) and Cx. (Phc.). Culex (Cux.) chidesteri, Cx. (Cux.) coronator, Cx. (Cux.) interrogator, Cx. (Cux.) quinquefasciatus, Cx. (Cux.) nigripalpus/Cx. (Cux.) thriambus, and Cx. (Phc.) lactator were identified directly with a multiplexed primer cocktail comprising a conserved forward primer and specific reverse primers targeting ribosomal DNA (rDNA). Culex nigripalpus and Cx. thriambus were differentiated by restriction digest of homologous amplicons. The assay was developed and optimized using well-characterized specimens from Guatemala and the United States and field tested with unknown material from Guatemala. This assay will be a valuable tool for mosquito identification in entomological and arbovirus ecology studies in Guatemala. PMID:20682869

Objective To determine lead level primarily in Culex quinquefasciatus (Cx. quinquefasciatus), and Culex gelidus (Cx. gelidus) larvae inhabiting lead consuming factories, and to putatively estimate eco-toxicological impact of effluents from the firms. Methods Third instars larvae were sampled by standard dipping method and lead concentrations in the larvae and their respective surrounding factory aquatic environments were determined through standard atomic absorption spectrophotometry (AAS). Results Cx. quinquefasciatus was the most abundant species followed by Cx. gelidus. The levels of lead were higher in the Cx. quinquefasciatus (1.08-47.47 µg/g), than in the wastewaters surface (0.01-0.78 µg/mL) from the factories or closer areas around factories. Other species were not reaching the criteria for lead determination. Conclusions The Cx. quinquefasciatus larvae can bio-accumulate the metal and can potentially serve as a biomarker of lead contamination, to complemente conventional techniques. PMID:23569727

Mitochondrial DNA sequence data from the control region and 12S rRNA in leopard frogs from the Sierra El Aguaje of southern Sonora, Mexico, together with GenBank sequences, were used to infer taxonomic identity and provide phylogenetic hypotheses for relationships with other members of the Rana pipiens complex. We show that frogs from the Sierra El Aguaje belong to the Rana berlandieri subgroup, or Scurrilirana clade, of the R. pipiens group, and are most closely related to Rana magnaocularis from Nayarit, Mexico. We also provide further evidence that Rana magnaocularis and R. yavapaiensis are close relatives.

Culex bidens and C. interfor, implicated in arbovirus transmission in Argentina, are sister species, only distinguishable by feature of the male genitalia; however, intermediate specimens of the species in sympatry have been found. Fourth-instar larvae and females of both species share apomorphic features, and this lack of clear distinction creates problems for specific identification. Geometric morphometric traits of these life stages also do not distinguish the species. The aim of the present study was to assess the taxonomic status of C. bidens and C. interfor using two mitochondrial genes and to determine the degree of their reproductive isolation using microsatellite loci. Sequences of the ND4 and COI genes were concatenated in a matrix of 993 nucleotides and used for phylogenetic and distance analyses. Bayesian and maximum parsimony inferences showed a well resolved and supported topology, enclosing sequences of individuals of C. bidens (0.83 BPP, 73 BSV) and C. interfor (0.98 BPP, 97 BSV) in a strong sister relationship. The mean K2P distance within C. bidens and C. interfor was 0.3% and 0.2%, respectively, and the interspecific variation was 2.3%. Bayesian clustering also showed two distinct mitochondrial lineages. All sequenced mosquitoes were successfully identified in accordance with the best close match algorithm. The low genetic distance values obtained indicate that the species diverged quite recently. Most morphologically intermediate specimens of C. bidens from Córdoba were heterozygous for the microsatellite locus GT51; the significant heterozygote excess observed suggests incomplete reproductive isolation. However, C. bidens and C. interfor should be considered good species: the ventral arm of the phallosome of the male genitalia and the ND4 and COI sequences are diagnostic characters. PMID:28235083

Culex bidens and C. interfor, implicated in arbovirus transmission in Argentina, are sister species, only distinguishable by feature of the male genitalia; however, intermediate specimens of the species in sympatry have been found. Fourth-instar larvae and females of both species share apomorphic features, and this lack of clear distinction creates problems for specific identification. Geometric morphometric traits of these life stages also do not distinguish the species. The aim of the present study was to assess the taxonomic status of C. bidens and C. interfor using two mitochondrial genes and to determine the degree of their reproductive isolation using microsatellite loci. Sequences of the ND4 and COI genes were concatenated in a matrix of 993 nucleotides and used for phylogenetic and distance analyses. Bayesian and maximum parsimony inferences showed a well resolved and supported topology, enclosing sequences of individuals of C. bidens (0.83 BPP, 73 BSV) and C. interfor (0.98 BPP, 97 BSV) in a strong sister relationship. The mean K2P distance within C. bidens and C. interfor was 0.3% and 0.2%, respectively, and the interspecific variation was 2.3%. Bayesian clustering also showed two distinct mitochondrial lineages. All sequenced mosquitoes were successfully identified in accordance with the best close match algorithm. The low genetic distance values obtained indicate that the species diverged quite recently. Most morphologically intermediate specimens of C. bidens from Córdoba were heterozygous for the microsatellite locus GT51; the significant heterozygote excess observed suggests incomplete reproductive isolation. However, C. bidens and C. interfor should be considered good species: the ventral arm of the phallosome of the male genitalia and the ND4 and COI sequences are diagnostic characters.

Culex (Melanoconion) pedroi Sirivanakarn & Belkin 1980 and Culex (Melanoconion) ribeirensis Forattini & Sallum 1985 are two morphologically very similar species of the Pedroi subgroup of mosquitoes in the Spissipes section of the subgenus Melanoconion of the genus Culex L. 1758. We carried out an analysis of the mitochondrial cytochrome c oxidase I (COI) DNA variation between these two species. The recent observation of sympatric coexistence in a forested area of Rio de Janeiro State (Brazil) triggered the need to validate these two species previously identified based on morphology. We concluded that the COI is a useful tool for identification of Cx. pedroi and Cx. ribeirensis.

Culex mosquitoes feed on a wide range of nectars consisting of mostly carbohydrates and amino acids, however, little is known about the utilization and effects of these different carbohydrates and their accompanying amino acids on longevity. Culex quinquefasciatus larvae were reared on high and low...

Early development in the frog model, Xenopus laevis, is governed by RNAs, localized to the vegetal cortex of the oocyte. These RNAs include Xdazl RNA, which is involved in primordial germ cell formation, and VegT RNA, which specifies the mesoderm and endoderm. In order to determine whether orthologues of these RNAs are localized and have similar functions in other frogs, we cloned RpDazl and RpVegT from Rana pipiens, a frog that is phylogenetically distant from X. laevis. RNAs from both genes are localized to the vegetal cortex of the R. pipiens oocyte, indicating that the vegetal localization is likely the basal state. The animal location of EcVegT RNA in Eleutherodactylus coqui that we found previously (Beckham et al., 2003) is then a derived state, probably due to the great increase in egg size required for direct development of this species. To answer the question of function, we injected RpVegT or EcVegT RNAs into X. laevis embryos, and assayed animal caps for gene expression. Both of these RNAs induced the expression of endodermal, mesodermal, and organizer genes, showing that the function of RpVegT and EcVegT as meso-endodermal determinants is conserved in frogs. The RNA localizations and the function of VegT orthologues in germ layer specification may be synapomorphies for anuran amphibians.

As a part of studies on the reproduction of laboratory maintained frogs, wild-caught Rana pipiens were ovulated and maintained at 22-27 degrees C for up to 18 months. Vitellogenic oocytes were periodically staged and counted, and a "maturity index" was calculated to assess the progress of the vitellogenic cycle. The initial cycle was similar to that of wild frogs except that the first oocytes to reach stage 5 (mature eggs) usually began to degenerate before later starting oocytes became mature. In addition, a second cycle began before the first was completed. After more than 1 year at room temperature, abnormal cycles were common. Ovaries of such animals contained very few mature eggs. Many of their oocytes were in early stages of vitellogenesis or, if pigmented, had begun to degenerate. These deficiencies were partially corrected in females placed in 4 degrees C for 4-6 weeks. The average number of mature eggs increased 15-fold and ovary weights more than doubled. Oviduct weights almost doubled. Although the rates of cooling, photoperiod, and nutritional status could be important influences, the results imply that cold treatment alone increases estrogen secretion. We suggest that low estrogen secretion may account for the reproductive deficiencies seen in R. pipiens cultured at room temperature.

The authors study the sensitive potential evoked from point of Erb and from cervical spine in C6-C7, obtained by stimulation of median nerve in a control group (normals) and in a greater group of 40 cases from patients affected by radiculopathie with or without discal protrusion and by myelopathie spondiloartrosic. The date supply significant informations and are (obicurred in analytique) analyzed with accuracy.

The sense of "Presence" (evolving from "telepresence") has always been associated with virtual reality research and is still an exceptionally mystifying constituent. Now the study of presence clearly spans over various disciplines associated with cognition. This paper attempts to put forth a concept that argues that it's an experience of an "Evoked Reality (ER)" (illusion of reality) that triggers an "Evoked Presence (EP)" (sense of presence) in our minds. A Three Pole Reality Model is proposed to explain this phenomenon. The poles range from Dream Reality to Simulated Reality with Primary (Physical) Reality at the center. To demonstrate the relationship between ER and EP, a Reality-Presence Map is developed. We believe that this concept of ER and the proposed model may have significant applications in the study of presence, and in exploring the possibilities of not just virtual reality but also what we call "reality."

Among members of the order Mononegavirales, RNA splicing events have been found only in the family Bornaviridae. Here, we report that a new rhabdovirus isolated from the mosquito Culex tritaeniorhynchus replicates in the nuclei of infected cells and requires RNA splicing for viral mRNA maturation. The virus, designated Culex tritaeniorhynchus rhabdovirus (CTRV), shares a similar genome organization with other rhabdoviruses, except for the presence of a putative intron in the coding region for the L protein. Molecular phylogenetic studies indicated that CTRV belongs to the family Rhabdoviridae, but it is yet to be assigned a genus. Electron microscopic analysis revealed that the CTRV virion is extremely elongated, unlike virions of rhabdoviruses, which are generally bullet shaped. Northern hybridization confirmed that a large transcript (approximately 6,500 nucleotides [nt]) from the CTRV L gene was present in the infected cells. Strand-specific reverse transcription-PCR (RT-PCR) analyses identified the intron-exon boundaries and the 76-nt intron sequence, which contains the typical motif for eukaryotic spliceosomal intron-splice donor/acceptor sites (GU-AG), a predicted branch point, and a polypyrimidine tract. In situ hybridization exhibited that viral RNAs are primarily localized in the nucleus of infected cells, indicating that CTRV replicates in the nucleus and is allowed to utilize the host's nuclear splicing machinery. This is the first report of RNA splicing among the members of the family Rhabdoviridae.

The increase in ultraviolet light intensity levels due to ozone depletion recently has been linked to the decline in amphibian population. In this experiment, eggs and larvae of Rana pipiens were subjected to differing amounts of ultraviolet radiation to determine the effects of ultraviolet light on the development of amphibian tadpoles. The total length, length of body without tail, and maximum width of each specimen was recorded for a month of the tadpoles` development, including several measurements after the ultraviolet exposures were concluded. It was found that ultraviolet exposure significantly reduced the size of the organisms in comparison with the control group in all three measured areas. Ultraviolet radiation altered the health and appearance of the exposed organisms and was lethal at large amounts. This experiment showed that ultraviolet radiation could cause many problems in developing amphibians. By slowing their development and physically weakening predation, thus contributing to a decline in overall population levels.

Rana pipiens eggs dividing normally in diluted Ringer's solution show an increase in transmembrane potential inside negative, a decrease in resistance, and no change in total surface membrane capacitance at the appearance of a division furrow. Furrows of eggs in solutions with the tonicity of full Ringer develop partially, then regress so that the surface is again spherical. The potential and resistance changes are greater and substantial increases in capacitance occur when furrowing is so inhibited. It is proposed that the electrical changes at division are due to the introduction of new plasma membrane, between the blastomeres, having selective permeability to K and a low resistance compared to the outer spherical membrane. A narrow gap between blastomeres limits current flow through new membrane during normal division. A direct exposure of new membrane to the bathing medium when furrowing is disrupted results in larger changes in potential and resistance and permits the capacitance of new membrane to be detected. PMID:5691712

Culex (Culiciomyia) spathifurca (Edwards) is recorded for the 1st time in India. Immature specimens of this species were collected from tree holes in a mangrove forest. Notes on the larval habitat are given.

Culex mosquitoes are important vectors of pathogens and parasites causing diseases such as West Nile virus, St. Louis encephalitis, Japanese encephalitis, Venezuelan equine encephalitis and bancroftian filariasis. Surveillance of these species is based on traps using conventional mosquito attractan...

The sense of “Presence” (evolving from “telepresence”) has always been associated with virtual reality research and is still an exceptionally mystifying constituent. Now the study of presence clearly spans over various disciplines associated with cognition. This paper attempts to put forth a concept that argues that it’s an experience of an “Evoked Reality (ER)” (illusion of reality) that triggers an “Evoked Presence (EP)” (sense of presence) in our minds. A Three Pole Reality Model is proposed to explain this phenomenon. The poles range from Dream Reality to Simulated Reality with Primary (Physical) Reality at the center. To demonstrate the relationship between ER and EP, a Reality-Presence Map is developed. We believe that this concept of ER and the proposed model may have significant applications in the study of presence, and in exploring the possibilities of not just virtual reality but also what we call “reality.” PMID:23550234

Electrical evoked otoacoustic emissions (EEOAE) are used to investigate in vivo cochlear electromechanical function. Round window electrical stimulation gives rise to a broad frequency EEOAE response, from 100 Hz or below to 40 kHz in guinea pigs. Placing bipolar electrodes very close to the basilar membrane (in the scala vestibuli and scala tympani) gives rise to a much narrower frequency range of EEOAE, limited to around 20 kHz when the electrodes are placed near the 18 kHz best frequency place. Model predictions using a three dimensional fluid model in conjunction with a simple model for outer hair cell (OHC) activity are used to interpret the experimental results. The model is solved using a 2.5D finite-element formulation. Predictions show that the high-frequency limit of the excitation is determined by the spatial extent of the current stimulus (also called the current spread). The global peaks in the EEOAE spectra are interpreted as constructive interference between electrically evoked backward traveling waves and forward traveling waves reflected from the stapes. Steady-state response predictions of the model are presented.

Time-variant muscle responses under electrical stimulation (ES) are often problematic for all the applications of neuroprosthetic muscle control. This situation limits the range of ES usage in relevant areas, mainly due to muscle fatigue and also to changes in stimulation electrode contact conditions, especially in transcutaneous ES. Surface electrodes are still the most widely used in noninvasive applications. Electrical field variations caused by changes in the stimulation contact condition markedly affect the resulting total muscle activation levels. Fatigue phenomena under functional electrical stimulation (FES) are also well known source of time-varying characteristics coming from muscle response under ES. Therefore, it is essential to monitor the actual muscle state and assess the expected muscle response by ES so as to improve the current ES system in favor of adaptive muscle-response-aware FES control. To deal with this issue, we have been studying a novel control technique using evoked electromyography (eEMG) signals to compensate for these muscle time-variances under ES for stable neuroprosthetic muscle control. In this perspective article, I overview the background of this topic and highlight important points to be aware of when using ES to induce the desired muscle activation regardless of the time-variance. I also demonstrate how to deal with the common critical problem of ES to move toward robust neuroprosthetic muscle control with the Evoked Electromyographically Controlled Electrical Stimulation paradigm. PMID:27471448

Background The Coronator Group encompasses Culex coronator Dyar & Knab, Culex camposi Dyar, Culex covagarciai Forattini, Culex ousqua Dyar, Culex usquatissimus Dyar, Culex usquatus Dyar and Culex yojoae Strickman. Culex coronator has the largest geographic distribution, occurring in North, Central and South America. Moreover, it is a potential vector-borne mosquito species because females have been found naturally infected with several arboviruses, i.e., Saint Louis Encephalitis Virus, Venezuelan Equine Encephalitis Virus and West Nile Virus. Considering the epidemiological importance of Cx. coronator, we investigated the wing shape diversity of Cx. coronator from South and Southeast Brazil, a method to preliminarily estimate population diversity. Methods Field-collected immature stages of seven populations from a large geographical area in Brazil were maintained in the laboratory to obtain both females and males linked with pupal and/or larval exuviae. For each individual female, 18 landmarks of left wings were marked and digitalized. After Procrustes superimposition, discriminant analysis of shape was employed to quantify wing shape variation among populations. The isometric estimator centroid size was calculated to assess the overall wing size and allometry. Results Wing shape was polymorphic among populations of Cx. coronator. However, dissimilarities among populations were higher than those observed within each population, suggesting populational differentiation in Cx. coronator. Morphological distances between populations were not correlated to geographical distances, indicating that other factors may act on wing shape and thus, determining microevolutionary patterns in Cx. coronator. Despite the population differentiation, intrapopulational wing shape variability was equivalent among all seven populations. Conclusion The wing variability found in Cx. coronator populations brings to light a new biological problem to be investigated: the population genetics of

When late 3rd or early 4th-instar larvae of Culex tarsalis Coquillett and Culex quinquefasciatus Say mosquitoes were treated with sublethal dosages of neem insecticide until pupation, the blood-feeding activity of the resulting adults was essentially the same as that of untreated controls. In contrast, blood-feeding activity was suppressed when newly emerged adults were fed continuously on 10 parts per million (ppm) or 50 ppm azadirachtin (AZ) in 10% sucrose solution for seven days. Fecundity was also reduced by the various neem treatments. When late 3rd or early 4th-instar larvae were treated with 0.010 ppm AZ to pupation, the resultant females had a lower rate of oviposition than did the untreated controls after a full blood meal. When late instar larvae were treated at 0.005 ppm and 0.010 ppm AZ, the resultant females produced smaller egg rafts after a full blood meal, as compared to the controls, but egg viability was not affected. In newly emerged adults feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose for seven days (before blood feeding), the oviposition rate, size of egg raft, and hatching rate of the eggs after a full blood meal were all reduced. When newly blood-fed adults were fed continuously on 10 ppm and 50 ppm AZ in 10% sucrose for five days, their oviposition rate was lower than controls in most cases, but the egg raft size and viability of eggs were not affected. In freshly blood-fed females topically treated with AZ with 1 or 5 micrograms/female, the oviposition rate and size of egg rafts were generally reduced. The females receiving topical treatment laid eggs and their hatching was not affected. The longevity of adult females feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose solution after emergence was reduced, whereas, the longevity of males was only affected at the higher concentration.

Figures are presented of computer-averaged auditory evoked responses (AERs) that point to the existence of a completely endogenous brain event. A series of regular clicks or tones was administered to the ear, and 'odd-balls' of different intensity or frequency respectively were included. Subjects were asked either to ignore the sounds (to read or do something else) or to attend to the stimuli. When they listened and counted the odd-balls, a P3 wave occurred at 300msec after stimulus. When the odd-balls consisted of omitted clicks or tone bursts, a similar response was observed. This could not have come from auditory nerve, but only from cortex. It is evidence of recognition, a conscious process.

Three male, wild-caught northern leopard frogs (Rana pipiens) died over a 1-week period with no previous history of clinical illness or disease. Noteworthy necropsy findings in one of the three frogs included depleted fat bodies in the coelomic cavity, indicating a poor nutritional condition, and a heavy parasite burden in the lungs. The location of infection and morphologic characteristics of the parasite were consistent with infection by the common lung fluke, Haematoloechus sp. In contrast to the heavy fluke load, only minor microscopic changes were observed in the lungs. Lesions included mild hypertrophy of the bronchiolar epithelium, with few submucosal inflammatory cells consisting predominantly of lymphocytes. Subsequent review of the literature revealed little about the pathologic effects of these parasites except that small numbers are thought to cause the host little harm. Our findings suggest that even with a large number of parasites, there is minimal pathologic impact in the lungs. We conclude that heavy lung-fluke infection should not be diagnosed as the sole or major etiology of death or illness in leopard frogs.

The analyses of necrophagous insects feeding on a corpse can be successfully used to estimate the minimum time since death. A minimum time frame is sometimes an underestimate, but it is actually the only method that can provide such information when decomposed remains are found at a crime scene. Many insects are known to be colonisers of a corpse, but because there is an endless spectrum of crime scene environments, the development data bases for necrophagous insects is incomplete. The two cases detailed in this paper show different entomological patterns due to the different environments (well and burial) and locations (south and central Italy) where the two cadavers were found. Common to both of these cases' was the discovery of the corpse in the same period of the year (January) and the presence of Syritta pipiens (Diptera: Syrphidae), a species that has never been associated with deceased humans. The ecological information concerning this insect was used in combination with the more typical entomofauna found on the corpse to provide a minimum post mortem interval.

The security of the northern leopard frog (Rana pipiens) varies spatially with populations east and west of North Dakota considered as secure and at risk, respectively. We used genetic markers to characterize the conservation status of northern leopard frog populations across North Dakota. We used multiple regression analyses and model selection to evaluate correlations of expected heterozygosity (HE) with the direct and additive effects of: i) geographic location,ii) wetland density and iii) average annual precipitation. There was lower genetic diversity in the western portion of the state due to lower levels of diversity for populations southwest of the Missouri River. This may reflect a refugial/colonization signature for the only non-glaciated area of North Dakota. Genetic diversity was also positively associated with wetland densities which is consistent with the reliance of this species on a mosaic of wetlands. Our findings suggest that populations in the southwestern part of North Dakota are of higher conservation concern, a finding consistent with the higher risk noted for northern leopard frog populations in most states west of North Dakota. Our findings also pose the hypothesis that climate change induced changes in wetland densities will reduce genetic diversity of northern leopard frog populations.

1. Responses of enzymic characteristics of gastrocnemius muscle were studied when frogs (Rana pipiens) were exposed to cold environment (4 degrees C). 2. The content of adenosine triphosphate (ATP) decreased significantly after cold exposure. This decrease was greater in starved than in fed frogs. 3. Although the glycogen content did not change, lactate levels were lower in cold-exposed than room-temperature (control) frogs. No change was observed in glycogen and lactate between fed and unfed frogs kept at 4 degrees C for 2 months. Lactate dehydrogenase activity tended to increase during chronic cold exposure, but not significantly. 4. The activities of citrate synthase, cytochrome oxidase, and beta-hydroxyacyl CoA dehydrogenase were higher in gastrocnemius of chronically cold-exposed frogs than in room-temperature controls. This increase was statistically significant only in the muscles of starved frogs; these muscles had the greatest decrease in ATP. 5. It was suggested that chronic cold exposure decreases skeletal muscle ATP content but may not affect glycolysis. The data also suggested that the decrease in ATP content stimulates mitochondrial biogenesis which increases enzyme activities. PMID:3261790

The management of neonatal hyperbilirubinemia is very standardized. However, there is a lack of an objective method to evaluate the cerebral effects of bilirubin apart from brainstem auditory evoked potentials. There were few studies evaluating the effects of hyperbilirubinemia or phototherapy on the visual pathway in infants with hyperbilirubinemia. Serial visual evoked potentials of two groups of term neonates (N = 24)--group 1 with moderate hyperbilirubinemia (n = 16) and group 2 with severe hyperbilirubinemia (n = 8)--were evaluated prospectively. All infants had regular physical, neurologic, visual, and auditory evaluations until 3 years. Four (16%) had abnormal visual evoked potentials before 1 year, and the abnormalities returned to normal thereafter. There was no significant difference in visual evoked potentials between the two groups. All had normal neurodevelopmental status by 3 years, with the exception of one child from the severe group with ABO incompatibility with transient mild motor delay, hypotonia, and abnormal visual evoked potential. There were no abnormal effects of phototherapy on visual evoked potentials in infants with neonatal hyperbilirubinemia after 1 year of age. Although our sample size was small, the results suggest that the effects of hyperbilirubinemia on visual evoked potentials might be transient. (J Child Neurol 2006;21:58-62).

Background Zanzibar has a long history of lymphatic filariasis (LF) caused by the filarial parasite Wuchereria bancrofti, and transmitted by the mosquito Culex quinquefasciatus Say. The LF Programme in Zanzibar has successfully implemented mass drug administration (MDA) to interrupt transmission, and is now in the elimination phase. Monitoring infections in mosquitoes, and assessing the potential role of interventions such as vector control, is important in case the disease re-emerges as a public health problem. Here, we examine Culex mosquito species from the two main islands to detect W. bancrofti infection and to determine levels of susceptibility to the insecticides used for vector control. Methods Culex mosquitoes collected during routine catches in Vitongoji, Pemba Island, and Makadara, Unguja Island were tested for W. bancrofti infection using PCR. Insecticide bioassays on Culex mosquitoes were performed to determine susceptibility to permethrin, deltamethrin, lambda-cyhalothrin, DDT and bendiocarb. Additional synergism assays with piperonyl butoxide (PBO) were used for lambda-cyhalothrin. Pyrosequencing was used to determine the kdr genotype and sequencing of the mitochondrial cytochrome oxidase I (mtCOI) subunit performed to identify ambiguous Culex species. Results None of the wild-caught Culex mosquitoes analysed were found to be positive for W. bancrofti. High frequencies of resistance to all insecticides were found in Wete, Pemba Island, whereas Culex from the nearby site of Tibirinzi (Pemba) and in Kilimani, Unguja Island remained relatively susceptible. Species identification confirmed that mosquitoes from Wete were Culex quinquefasciatus. The majority of the Culex collected from Tibirinzi and all from Kilimani could not be identified to species by molecular assays. Two alternative kdr alleles, both resulting in a L1014F substitution were detected in Cx. quinquefasciatus from Wete with no homozygote susceptible detected. Metabolic resistance to

Background. Objective electrophysiological methods for investigations of the organ of smell consist in recordings of olfactory cortex responses to specific, time restricted odor stimuli. In hypothyroidism have impaired sense of smell. Material and Methods. Two groups: control of 31 healthy subjects and study group of 21 with hypothyroidism. The inclusion criterion for the study group was the TSH range from 3.54 to 110 μIU/mL. Aim. Assessment of the latency time of evoked responses from the olfactory nerve N1 and the trigeminal nerve N5 using two smells of mint and anise in hypothyroidism. Results. The smell perception in subjective olfactory tests was normal in 85% of the hypothyroid group. Differences were noticed in the objective tests. The detailed intergroup analysis of latency times of recorded cortical responses PN5 and PN1 performed by means between the groups of patients with overt clinical hypothyroidism versus subclinical hypothyroidism demonstrated a significant difference (p < 0.05) whereas no such differences were found between the control group versus subclinical hypothyroidism group (p > 0.05). Conclusion. We can conclude that registration of cortex potentials at irritation of olfactory and trigeminal nerves offers possibilities for using this method as an objective indicator of hypothyroidism severity and prognostic process factor.

Background. Objective electrophysiological methods for investigations of the organ of smell consist in recordings of olfactory cortex responses to specific, time restricted odor stimuli. In hypothyroidism have impaired sense of smell. Material and Methods. Two groups: control of 31 healthy subjects and study group of 21 with hypothyroidism. The inclusion criterion for the study group was the TSH range from 3.54 to 110 μIU/mL. Aim. Assessment of the latency time of evoked responses from the olfactory nerve N1 and the trigeminal nerve N5 using two smells of mint and anise in hypothyroidism. Results. The smell perception in subjective olfactory tests was normal in 85% of the hypothyroid group. Differences were noticed in the objective tests. The detailed intergroup analysis of latency times of recorded cortical responses PN5 and PN1 performed by means between the groups of patients with overt clinical hypothyroidism versus subclinical hypothyroidism demonstrated a significant difference (p < 0.05) whereas no such differences were found between the control group versus subclinical hypothyroidism group (p > 0.05). Conclusion. We can conclude that registration of cortex potentials at irritation of olfactory and trigeminal nerves offers possibilities for using this method as an objective indicator of hypothyroidism severity and prognostic process factor. PMID:27656655

This study was conducted to isolate and identify natural entomopathogenic fungi from female Culex quinquefasciatus and to test their adulticidal activity. Field-collected female C. quinquefasciatus died early and were placed on a Saboraud's dextrose agar plates for growth and isolation of natural entomopathogenic fungi. The plates were maintained in an incubator at 24+/-2 degrees C for 3 days. Four fungal species were isolated in two genera namely, Aspergillus and Fusarium. The identified fungal species were A. niger, A. flavus, A. nidulans var acristatus (ITCC-6327.04), and F. pallidoroseum (ITCC-6324.06). Adult bioassays were carried out using spore-impregnated paper in WHO-holding tubes. F. pallidoroseum was found to be more effective than the others. Exposure of C. quinquefasciatus to spores of A. flavus and A. niger for 4 h caused 5.53% and 5.51% mortality in the mosquitoes within a week, respectively. All the female C. quinquefasciatus were killed within 4 days of exposure to F. pallidoroseum at a concentration of 1.11 x 10(10) conidia per m2. Significant difference of longevity was observed between the F. pallidoroseum-treated C. quinquefasciatus and control mosquitoes. The LT50 of F. pallidoroseum was 2.08 days for 4 h exposure to C. quinquefasciatus. Results of the present study confirm that F. pallidoroseum is one of the alternative biological control agents of adult mosquitoes.

Aedes aegypti and Culex quinquefasciatus were found in large numbers emerging from septic tanks in southern Puerto Rico during the dry season. Previous studies suggested that Ae. aegypti uses subterranean aquatic habitats only during dry periods when surface containers do not have water. This research investigated whether septic tanks are alternative aquatic habitats that this mosquito uses during unfavorable times of the year, or whether Ae. aegypti uses this aquatic habitat throughout the year. To assess temporal change, exit traps were used to collect mosquitoes emerging from septic tanks in Playa/Playita, southern Puerto Rico, from November 2006 to October 2007. We also investigated the hypotheses that (1) the production of Ae. aegypti in septic tanks was larger than in surface containers and (2) adult mosquitoes emerging from septic tanks were larger than those emerging from surface containers. This study demonstrated that unsealed septic tanks produced large numbers of Ae. aegypti and Cx. quinquefasciatus throughout the year, without any significant relationship with rainfall. The number of adult Ae. aegypti emerging per day from septic tanks in each community was 3 to 9 times larger than those produced in surface containers. It was also demonstrated that Ae. aegypti emerging from septic tanks were significantly larger than those emerging from surface container habitats. It is recommended that dengue prevention programs include regular inspection and maintenance of septic tanks in communities lacking sewerage.

The dynamics of a mosquito population depends heavily on climatic variables such as temperature and precipitation. Since climate change models predict that global warming will impact on the frequency and intensity of rainfall, it is important to understand how these variables affect the mosquito populations. We present a model of the dynamics of a Culex quinquefasciatus mosquito population that incorporates the effect of rainfall and use it to study the influence of the number of rainy days and the mean monthly precipitation on the maximum yearly abundance of mosquitoes Mmax. Additionally, using a fracturing process, we investigate the influence of the variability in daily rainfall on Mmax. We find that, given a constant value of monthly precipitation, there is an optimum number of rainy days for which Mmax is a maximum. On the other hand, we show that increasing daily rainfall variability reduces the dependence of Mmax on the number of rainy days, leading also to a higher abundance of mosquitoes for the case of low mean monthly precipitation. Finally, we explore the effect of the rainfall in the months preceding the wettest season, and we obtain that a regimen with high precipitations throughout the year and a higher variability tends to advance slightly the time at which the peak mosquito abundance occurs, but could significantly change the total mosquito abundance in a year.

Alpha-terthienyl is toxic to late embryonic stages ofRana pipiens in the presence of sunlight. Neither α-terthienyl alone in the dark nor a previously photolyzed solution of α-terthienyl has comparable activity. The LC50 was 0.11 ppm with 30 min of exposure and 0.018 ppm with 2 hr of exposure to sunlight. Anthracene, a representative example of polycyclic aromatic hydrocarbons widely distributed in the environment, also showed similar phototoxicity, with an LC50 of 0.065 ppm after 30 min of exposure and 0.025 ppm after 5 hr.

Bioinsecticides are shown to be useful in control programs to prevent several diseases, based on their specificity and efficiency against insect vectors. In the current study a bioinsecticide based on Bacillus sphaericus was produced using a white soybean culture medium and applied to larvae of Culex quinquefasciatus, the susceptible species, and Aedes aegypti, the refractory species used as the negative control. Efficacy was compared with that of the product fermented with the Luria Bertani (LB) reference medium. The experiments showed that C. quinquefasciatus was highly susceptible to the product prepared with white soybean meal, reaching 100% larval mortality even at 10mg/L, while A. aegypti failed to reach 70% mortality at a concentration of 1g/L. By comparison with the reference medium, the proposed culture medium showed high larvicidal power, reaching a LD90 of 2.26 mg/L, while 4.37 mg/L was needed for the LB medium to achieve the same mortality rate. Cost comparison between the formulations favored the use of the bioinsecticide produced with white soybean meal. After factoring in the LD90 value, the cost ratio favored the new raw material by nearly 1:220.

Introduction of irrigation projects in developing nations has often been blamed for aggravating the problem of mosquito-borne diseases by creating ideal larval habitats for vector mosquitoes. However, whereas several studies have demonstrated the relationship between malaria vectors and irrigation, little work has been done on culicine mosquitoes despite their potential in transmission of filariasis and arboviruses and their significant biting nuisance in these areas. This study examined the diversity of Culex mosquito fauna and their larval habitats at two sites (Murinduko and Kiamachiri) in Mwea, Kenya over a 12-month period. The habitat types present at each site within a 200-meter radius around the study village, including randomly selected paddies and canals, were sampled every two weeks to examine the relationship between vegetation cover, water depth, turbidity, and Culex larval counts. Ten culicine species belonging to four genera were identified, with 73.1% of the total collection comprising of Culex duttoni and Cx. quinquefasciatus. Other species collected included Cx. annulioris, Cx. poicilipes, Cx. cinereus, Cx. tigripes, Cx. trifilatus, Aedes spp., Coquilettidia fuscopennata, and Ficalbia splendens. Murinduko was more diverse than Kiamachiri in terms of species richness (10 versus 7 species) and larval habitat diversity (11 versus 8 habitat types). Paddies, canals, and rain pools were the most diverse habitats in terms of species richness, and ditches, rock pools, and tree holes were the least diverse. Principal component and correlation analyses showed a strong association between three Culex species and the measured habitat characteristics. Culex poicilipes was strongly associated with floating vegetation, Cx. annulioris with clean water containing emergent vegetation, and Cx. quinquefasciatus was associated with turbid water. Seasonal changes in larval counts in water reservoirs and pool and ditch habitats were closely associated with rainfall

The emergence of several diseases affecting amphibian populations worldwide has prompted investigations into determinants of the occurrence and abundance of parasites in frogs. To understand the spatial scales and identify specific environmental factors that determine risks of parasitism in frogs, helminth communities in metamorphic frogs of the northern leopard frog (Rana pipiens) were examined in relation to wetland and landscape factors at local (1 km) and regional (10 km) spatial extents in an agricultural region of Minnesota (USA) using regression analyses, ordination, and variance partitioning techniques. Greater amounts of forested and woody wetland habitats, shorter distances between woody wetlands, and smaller-sized open water patches in surrounding landscapes were the most consistently positive correlates with the abundances, richness, and diversity of helminths found in the frogs. Wetland and local landscape variables were suggested as most important for larval trematode abundances, whereas local and regional landscape variables appeared most important for adult helminths. As previously reported, the sum concentration of atrazine and its metabolite desethylatrazine, was the strongest predictor of larval trematode communities. In this report, we highlight the additional influences of landscape factors. In particular, our data suggest that anthropogenic activities that have resulted in the loss of the availability and connectivity of suitable habitats in the surrounding landscapes of wetlands are associated with declines in helminth richness and abundance, but that alteration of wetland water quality through eutrophication or pesticide contamination may facilitate the transmission of certain parasite taxa when they are present at wetlands. Although additional research is needed to quantify the negative effects of parasitism on frog populations, efforts to reduce inputs of agrochemicals into wetlands to limit larval trematode infections may be warranted

Metomidate hydrochloride is an imidazole-based, nonbarbiturate hypnotic drug primarily used as an immersion sedation and anesthetic agent in freshwater and marine finfish. To the authors' knowledge, there is no documentation in the literature of its use in amphibians. In this study, 7 male and 4 female leopard frogs (Rana pipiens) were induced with metomidate hydrochloride via immersion bath at a concentration of 30 mg/L for 60 min. The pH of the induction solution ranged from 7.63 to 7.75. Each frog was then removed from the induction solution, rinsed, and recovered in 26.6 degrees C amphibian Ringer's solution. After 210 min in the Ringer's solution, the frogs were transferred to moist paper towels for recovery. Heart rate, gular and abdominal respiration rates, righting reflex, superficial and deep pain withdrawal reflexes, corneal and palpebral reflexes, and escape response were monitored and recorded at defined intervals during both induction and recovery. The average time to loss of righting reflex and escape response was 17.36 min and 17.82 min, respectively. Metomidate produced clinical sedation in all frogs (n = 11). Surgical anesthesia was achieved in only 27% (3/11), with an anesthetic duration that ranged from 9 to 20 min. Recovery times were extremely prolonged and varied, with a range from 313 min to longer than 600 min. The findings of this study indicate that metomidate hydrochloride is unsuitable as a sole anesthetic agent in leopard frogs, and further research is needed to evaluate its suitability in other amphibians.

Northern leopard frogs Rana pipiens exposed to PCB 126 (3,3',4,4',5-pentachlorobiphenyl) were examined for hepatic oxidative stress. In a dose-response study, northern leopard frogs were injected intraperitoneally with either PCB 126 in corn oil (0.2, 0.7, 2.3, or 7.8 mg/kg body weight) or corn oil alone. In a time-course study, frogs received 7.8 mg/kg or corn oil alone, and were examined at 1, 2, 3, and 4 wk after dosing. Hepatic concentrations of reduced glutathione (GSH), thiobarbituric acid-reactive substances (TBARS), and total sulfhydryls (total SH), as well as activities of glutathione peroxidase (GSH-P), GSSG reductase (GSSG-R), glucose-6-phosphate dehydrogenase (G-6-PDH), and glutathione S-transferase (GSH-S-T) were measured. In the dose-response experiment, few effects were apparent 1 wk after dosing. In the time-course experiment, significant changes were observed in the 7.8-mg/kg group at 2 wk or more posttreatment. Hepatic concentrations of GSH and TBARS were higher than in corresponding controls at wk 3 and 4; the activities of GSSG-R and GSH-S-T were higher than in controls at wk 2 and 4; and the activity of G-6-PDH was increased at wk 2 and 4. These data collectively indicate that altered glutathione metabolism and oxidative stress occurred and were indicative of both toxicity and induction of protective mechanisms in frogs exposed to PCB. A similar delay in response was reported in fish and may relate to lower metabolic rate and physiological reactions in ectothermic vertebrates

Northern leopard frogs (Rana [Lithobates] pipiens) are considered sensitive, threatened, or endangered in all western states and western Canadian provinces. Historically present in eastern Washington in 6 major river drainages, leopard frogs are now only known to occur at 2 localized areas in the Crab Creek drainage in Grant County. During the summers of 2002-2005, we surveyed both areas to document extent of leopard frog distributions and to describe habitat and vertebrate community characteristics associated with leopard frog site occupancy. At Gloyd Seeps, 2 juvenile leopard frogs were observed in a total of 8.2 person-days of searching along a 5-km stream reach. At Potholes Reservoir, we surveyed 243 wetland sites in 7 management units known to have been occupied by leopard frogs during the 1980s. We confirmed leopard frog presence at only 87 sites (36%) in 4 management units. Site occupancy models for individual ponds indicated that, compared to unoccupied sites, occupied sites had slightly greater pond depths, less tall emergent vegetation, more herbaceous vegetative cover, and fewer neighboring ponds containing nonnative predatory fish. Models developed at the 1-km2 scale indicated that occupied areas had greater average midsummer pond depths, fewer ponds occupied by bullfrogs (Rana [Lithobates] catesbeiana) and carp (Cyprinus carpio), and more herbaceous vegetation surrounding ponds. The Gloyd Seeps population now appears defunct, and the Potholes Reservoir population is in sharp decline. Unless management actions are taken to reduce nonnative fish and bullfrogs and to enhance wetland vegetation, leopard frogs may soon be extirpated from both sites and possibly, therefore, from Washington.

Understanding the transmission patterns of West Nile and St. Louis encephalitis viruses (family Flaviviridae, genus Flavivirus, WNV and SLEV) could result in an increased ability to predict transmission risk to humans. To examine transmission patterns between vector and host, we trapped mosquitoes in three Florida counties from June to November 2005 by using chicken-baited lard can mosquito traps. These traps were used to monitor for presence of WNV and SLEV in mosquitoes and subsequent transmission of these viruses to chickens. In total, 166,615 female mosquitoes were sorted into 4,009 pools based on species and bloodfed status, and they were tested for presence of WNV and SLEV. Sera from 209 chickens were tested for WNV and SLEV antibodies. We detected eight WNV-positive Culex nigripalpus Theobald mosquito pools; SLEV was not detected in any pools. Six positive pools were collected in August and September from Duval County, one pool in September from Manatee County, and one pool in November from Indian River County. Of the eight chickens potentially exposed to WNV, antibodies were detected in only one chicken, indicating a low rate of transmission relative to the observed mosquito infection rates. Low virus transmission rates relative to infection rates would suggest that using sentinel chicken seroconversion data as a means of arbovirus surveillance may underestimate the prevalence of WNV in the mosquito population. However, using mosquito infection rates may overestimate the risk of arboviral transmission. A variety of factors might account for the observed low level of transmission including a lack of viral dissemination in mosquito vectors. PMID:18533444

The mosquito Culex quinquefasciatus is a ubiquitous species that serves as a major vector for west nile virus and lymphatic filariasis. Ingestion of bloodmeal by females triggers a series of physiological processes in the midgut and also exposes them to infection by these pathogens. The bacteria normally harbored in the midgut are known to influence physiology and can also alter the response to various pathogens. The midgut bacteria in female Cx. quinquefasciatus mosquitoes collected over a large geographical area from India was studied. Examination of 16S ribosomal DNA amplicons from culturable microflora revealed the presence of 83 bacterial species belonging to 31 bacterial genera. All of these species belong to three phyla i.e. Proteobacteria, Firmicutes and Actinobacteria. Phylum Proteobacteria was the most dominant phylum (37 species), followed by Firmicutes (33 species) and Actinobacteria (13 species). Phylum Proteobacteria, was dominated by members of γ-proteobacteria class. The genus Staphylococcus was the largest genus represented by 11 species whereas Enterobacter was the most prevalent genus and recovered from all the field stations except Leh. Highest bacterial prevalence was observed from Bhuj (22 species) followed by Nagrota (18 species), Masimpur (18 species) and Hathigarh (16 species). Whereas, least species were observed from Leh (8 species). It has been observed that individual mosquito harbor extremely diverse gut bacteria and have very small overlap bacterial taxa in their gut. This variation in midgut microbiota may be one of the factors responsible for variation in disease transmission rates or vector competence within mosquito population. The present data strongly encourage further investigations to verify the potential role of the detected bacteria in mosquito for the transmission of lymphatic filariasis and west nile virus. To the best of our knowledge this is the first study on midgut microbiota of wild Cx. quinquefasciatus from over a

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) persistently infects many mosquito tissues, and it has been associated with cytopathological changes in midgut muscles and salivary glands. However, the effects of WNV infection on mosquito fitness (survival and reproduction) are not known. We conducted a life table study of individually housed female Culex tarsalis Coquillett. After an initial bloodmeal from a WNV-infected or uninfected chicken, mosquitoes were provided sucrose and offered weekly opportunities to feed on a hanging blood drop. WNV transmission status was determined by testing the remaining blood drop for virus after mosquito feeding. Dead mosquitoes and eggs were collected daily. Mosquito legs and bodies were tested for WNV, and eggs were counted and allowed to hatch. Two replicates of this experiment were performed, with a total of 62 mosquitoes that fed on a WNV-infected chicken (of which 21 became infected) and 43 mosquitoes that fed on an uninfected chicken. Fecundity of WNV-infected mosquitoes was significantly lower than that of uninfected mosquitoes, especially during the first oviposition. WNV infection was associated with smaller egg rafts, whereas increasing wing length and WNV titer in the legs had a positive effect on egg raft size. Additionally, infected mosquitoes had lower egg hatch rates than did uninfected mosquitoes. There were no significant differences in survival between infected and uninfected mosquitoes. Blood feeding rates were higher in infected mosquitoes than in uninfected mosquitoes. A small amount of virus (average, 378; range, 5-5000 plaque-forming units) was transmitted to the blood drops fed upon by infected mosquitoes. Although WNV infection negatively impacts mosquito reproduction, facets of mosquito biology that are critical to virus transmission success were either not affected (survival) or changed in such a way as to result in enhanced vectorial capacity (blood feeding).

Population growth and urbanization have increased the potential habitats, and consequently the abundance of Culex quinquefasciatus, the southern house mosquito, a vector of West Nile Virus in urban areas. Water quality is critical in larval habitat distribution and in providing microbial food resources for larvae. A mesocosm experiment was designed to demonstrate which specific components of water chemistry are conducive to larval Culex mosquitoes. Dose-response relationships between larval development and NO3 , NH4 , and PO4 concentrations in stream water were developed through this experiment to describe the isolated effects of each nutrient on pre-adult development. The emergence pattern of Culex mosquitoes was found to be strongly related to certain nutrients, and results showed that breeding sites with higher PO4 or NO3 concentrations had higher larval survival rates. High NO3 concentrations favor the development of male mosquitoes and suppress the development of female mosquitoes, but those adult females that do emerge develop faster in containers with high NO3 levels compared to the reference group. The addition of PO4 in the absence of nitrogen sources to the larval habitat slowed larval development, however, it took fewer days for larvae to reach the pupal stage in containers with combinations of NO3 and PO4 or NH4 and PO4 nutrients. Results from this study may bolster efforts to control WNV in urban landscapes by exploring water quality conditions of Culex larval habitats that produce adult mosquitoes.

Japanese encephalitis virus (JEV) is a flavivirus that is transmitted by Culex (Cx.) tritaeniorhynchus in tropical and subtropical regions of Asia. The endemic transmission cycle involves domestic pigs and avian species that serve as amplification hosts; humans are incidental hosts that cannot devel...

Fecundity of mosquitoes can vary with many factors, and can have a strong effect on population growth. This study reports the effects of body size, blood meal size and age on reproductive output of nulliparous Culex quinquefasciatus, a vector of arboviruses and other pathogens. Mated adult female m...

Magnesium is crucial for baculovirus transmission in Culex nigripalpus (Theobald) and Cx. quinquefasciatus (Say) larvae, both in the field and in the laboratory. However, the mechanistic role of magnesium in baculovirus transmission is unknown. To investigate the possible role of a host response fac...

Culex quinquefasciatus is one of the most important mosquito species in North America due to its status as a vector of arboviruses such as West Nile virus and its often close association with agricultural activities. A better understanding of the physiological attributes of these mosquitoes is crit...

Concentrations of permethrin and malathion found in droplets generated from ultra-low volume and low volume sprays used to control adult mosquito populations were evaluated for efficacy against Culex quinquefasciatus Say using a topical application bioassay. Although insecticide droplets will imping...

The role of lactic acid was evaluated for attraction of Culex nigripalpus, Culex quinquefasciatus, Culex tarsalis, and Aedes aegypti in the laboratory using a dual-port olfactometer. When lactic acid was combined with chicken odor, attraction was increased for Cx. quinquefasciatus compared to chicken odor alone but not for Cx. nigripalpus, Cx. tarsalis, and Ae. aegypti. Lactic acid combined with hand odor did not change attraction of Cx. tarsalis and Ae. aegypti but decreased attraction of Cx. nigripalpus and Cx. quinquefasciatus. The addition of lactic acid to CO(2) increased attraction of Ae. aegypti and Cx. quinquefasciatus but reduced attraction of Cx. nigripalpus and Cx. tarsalis. Use of commercial lactic acid baits with CO(2) resulted in a similar trend except for Cx. nigripalpus which showed no difference. A blend of lactic acid, acetone, and dimethyl disulfide was attractive to Ae. aegypti (63.4%) but elicited low responses by all Culex spp. (1.3-26.8%). Addition of the blend to CO(2) increased attraction of Ae. aegypti and Cx. quinquefasciatus but reduced attraction of Cx. nigripalpus and Cx. tarsalis. The mixture of compounds plus CO(2) was as attractive as a hand for Cx. quinquefasciatus, Cx. tarsalis, and Ae. aegypti.

Visual, somatosensory, and brainstem auditory evoked potentials provide functional quantitative assessment of the cerebral cortex and brainstem. Their contribution at the acute stage of coma concerns diagnosis, prognosis, and follow-up. Four patterns are observed in traumatic coma: pattern 1=dysfunction of the cerebral cortex, brainstem integrity: good prognosis in more than 80% of cases; pattern 2=midbrain dysfunction: prognosis depends on both the reversibility of midbrain dysfunction and the extent of associated axonal lesions in the hemispheric white matter; pattern 3=pontine dysfunction due to transtentorial herniation: ominous prognosis, this pattern must be early detected by continuous monitoring; pattern 4=brain death: we currently use evoked potentials at the only brain-death confirmatory test, even in sedated patients. The contribution of evoked potentials in vegetative or minimally responsive states concerns the identification of these patients whose state is determined by midbrain dysfunction and the evaluation of persisting cognitive abilities in individual cases.

In this report we describe the complete genome sequence of a nucleopolyhedrovirus that infects larval stages of the mosquito Culex nigripalpus (CuniNPV). The CuniNPV genome is a circular double-stranded DNA molecule of 108,252 bp and is predicted to contain 109 genes. Although 36 of these genes show homology to genes from other baculoviruses, their orientation and order exhibit little conservation relative to the genomes of lepidopteran baculoviruses. CuniNPV genes homologous to those from other baculoviruses include genes involved in early and late gene expression (lef-4, lef-5, lef-8, lef-9, vlf-1, and p47), DNA replication (lef-1, lef-2, helicase-1, and dna-pol), and structural functions (vp39, vp91, odv-ec27, odv-e56, p6.9, gp41, p74, and vp1054). Auxiliary genes include homologues of genes encoding the p35 antiapoptosis protein and a novel insulin binding-related protein. In contrast to these conserved genes, CuniNPV lacks apparent homologues of baculovirus genes essential (ie-1 and lef-3) or stimulatory (ie-2, lef-7, pe38) for DNA replication. Also, baculovirus genes essential or stimulatory for early-late (ie-1, ie-2), early (ie-0 and pe-38), and late (lef-6, lef-11, and pp31) gene transcription are not identifiable. In addition, CuniNPV lacks homologues of genes involved in the formation of virogenic stroma (pp31), nucleocapsid (orf1629, p87, and p24), envelope of occluded virions (odv-e25, odv-e66, odv-e18), and polyhedra (polyhedrin/granulin, p10, pp34, and fp25k). A homologue of gp64, a budded virus envelope fusion protein, was also absent, although a gene related to the other category of baculovirus budded virus envelope proteins, Ld130, was present. The absence of homologues of occlusion-derived virion (ODV) envelope proteins and occlusion body (OB) protein (polyhedrin) suggests that both CuniNPV ODV and OB may be structurally and compositionally different from those found in terrestrial lepidopteran hosts. The striking difference in genome

The distribution and abundance of emerging Culex spp. were assessed within narrow (width: 3 m) and wide (width: 20 m) bands of California bulrush (Schoenoplectus californicus) and in the open water adjacent to emergent vegetation in 2 marshes of an ammonia-dominated wastewater treatment wetland in southern California. Emerging mosquitoes were collected along transects perpendicular to the path of water flow at 3 distances (1.5, 5, and 10 m) from the vegetation-open water interface in the wide bands of emergent vegetation, at the center of narrow bands of emergent vegetation, and at 1.5 m from the edge of emergent vegetation in the open water. The width of vegetation bands (3 vs. 20 m) influenced the effectiveness of integrated mosquito management practices, especially the application of mosquito control agents. Mosquito production from the 2 marshes also differed up to 14-fold, suggesting that the distance between the shorelines (62 vs. 74 m) of each marsh also influenced the efficacy of mosquito control agents applied from the shore and boats. Hot spots of mosquito production (75424 female Culex/m2/day) were found within the wide bands of bulrush. During summer, the relative abundance of Culex stigmatosoma among emerging mosquitoes increased from the periphery to the center of wide bands of emergent vegetation. Culex erythrothorax emergence rates were comparatively similar among the transects in the wide bands of emergent vegetation. Culex tarsalis adults increased in number from the periphery to the center of wide bands of bulrush and, in May, were > 95% of emerged mosquitoes.

The usefulness of evoked potentials (EPs) in the diagnosis of multiple sclerosis is limited by its relatively low sensitivity to subclinical lesions. However, they are still a good tool to assess the integrity of afferent and efferent pathways and to quantify the severity of white matter involvement. Transversal and longitudinal studies have demonstrated good correlation between EP abnormalities and disability, suggesting that multimodal evoked potentials could be useful in monitoring the disease evolution in single patients and as surrogate end points in clinical trials.

Music is a universal feature of human societies, partly owing to its power to evoke strong emotions and influence moods. During the past decade, the investigation of the neural correlates of music-evoked emotions has been invaluable for the understanding of human emotion. Functional neuroimaging studies on music and emotion show that music can modulate activity in brain structures that are known to be crucially involved in emotion, such as the amygdala, nucleus accumbens, hypothalamus, hippocampus, insula, cingulate cortex and orbitofrontal cortex. The potential of music to modulate activity in these structures has important implications for the use of music in the treatment of psychiatric and neurological disorders.

The article reports on the first study of the evoked activity of the brain in epileptic patients (n = 20) following sleep deprivation. An analysis of the data obtained has revealed a tendency to the shortening of the peak latent intervals of visual evoked potentials in the range of 100-200 mu sec and the V component and the interpeak interval III-V of evoked auditory trunk potentials in patients with temporal epilepsy. The phenomenon may indicate the elimination of stabilizing control involving the specific conductive pathways and, possibly, an accelerated conduction of a specific sensor signal.

Diabetic neuropathy is a severe complication of long-standing diabetes and one of the major etiologies of neuropathic pain. Diabetes is associated with an increased formation of reactive oxygen species and the electrophilic dicarbonyl compound methylglyoxal (MG). Here we show that MG stimulates heterologously expressed TRPA1 in CHO cells and natively expressed TRPA1 in MDCK cells and DRG neurons. MG evokes [Ca(2+)]i-responses in TRPA1 expressing DRG neurons but is without effect in neurons cultured from Trpa1(-/-) mice. Consistent with a direct, intracellular action, we show that methylglyoxal is significantly more potent as a TRPA1 agonist when applied to the intracellular face of excised membrane patches than to intact cells. Local intraplantar administration of MG evokes a pain response in Trpa1(+/+) but not in Trpa1(-/-) mice. Furthermore, persistently increased MG levels achieved by two weeks pharmacological inhibition of glyoxalase-1 (GLO-1), the rate-limiting enzyme responsible for detoxification of MG, evokes a progressive and marked thermal (cold and heat) and mechanical hypersensitivity in wildtype but not in Trpa1(-/-) mice. Our results thus demonstrate that TRPA1 is required both for the acute pain response evoked by topical MG and for the long-lasting pronociceptive effects associated with elevated MG in vivo. In contrast to our observations in DRG neurons, MG evokes indistinguishable [Ca(2+)]i-responses in pancreatic β-cells cultured from Trpa1(+/+) and Trpa1(-/-) mice. In vivo, the TRPA1 antagonist HC030031 impairs glucose clearance in the glucose tolerance test both in Trpa1(+/+) and Trpa1(-/-) mice, indicating a non-TRPA1 mediated effect and suggesting that results obtained with this compound should be interpreted with caution. Our results show that TRPA1 is the principal target for MG in sensory neurons but not in pancreatic β-cells and that activation of TRPA1 by MG produces a painful neuropathy with the behavioral hallmarks of diabetic

Atrazine is the most commonly used herbicide in the United States and probably the world. Atrazine contamination is widespread and can be present in excess of 1.0 ppb even in precipitation and in areas where it is not used. In the current study, we showed that atrazine exposure (> or = to 0.1 ppb) resulted in retarded gonadal development (gonadal dysgenesis) and testicular oogenesis (hermaphroditism) in leopard frogs (Rana pipiens). Slower developing males even experienced oocyte growth (vitellogenesis). Furthermore, we observed gonadal dysgenesis and hermaphroditism in animals collected from atrazine-contaminated sites across the United States. These coordinated laboratory and field studies revealed the potential biological impact of atrazine contamination in the environment. Combined with reported similar effects in Xenopus laevis, the current data raise concern about the effects of atrazine on amphibians in general and the potential role of atrazine and other endocrine-disrupting pesticides in amphibian declines. PMID:12676617

Global amphibian declines suggest a major shift in the amount and quality of habitat for these sensitive taxa. Many species that were once widespread are now experiencing declines either in part of or across their historic range. The northern leopard frog (Rana [Lithobates] pipiens] has undergone significant declines particularly in the western United States and Canada. Leopard frog population losses in Nevada are largely due to habitat fragmentation and the introduction of nonnative fish, amphibian, and plant species. Only two populations remain in the Truckee and Carson River watersheds of western Nevada which represents the western boundary of this species range. We used sequence data for an 812 base pair fragment of the mitochondrial NADH dehydrogenase 1 (ND1) gene to support a native origin for western Nevada populations. All frogs had a single haplotype (W07) from the distinct western North America ND1 haplotype clade. Data from seven polymorphic microsatellite loci show that Truckee and Carson River populations are highly differentiated from each other and from leopard frogs collected from eastern Nevada sites. Lack of gene flow among and distinct color morphs among the western Nevada populations likely predates the current geographical isolation. Comparisons with other peripheral L. pipiens populations show western Nevada populations have similar levels of gene diversity despite their contemporary isolation (HE 0.411, 0.482). Restoration of leopard frog populations in these watersheds will be challenging given well-entrenched nonnative bullfrog populations and major changes to the riparian zone over the past century. Declines of once common amphibian species has become a major conservation concern. Contemporary isolation of populations on a species range periphery such as the leopard frog populations in the Truckee and Carson rivers further exacerbate extirpation risk as these populations are likely to have fewer genetic resources to adaptively respond to

The population genetics and phylogenetic relationships of Culex mosquitoes inhabiting the Sonoran Desert region of North America were studied using mitochondrial DNA and microsatellite molecular markers. Phylogenetic analyses of mitochondrial cytochrome c oxidase subunit I (COI) from mosquitoes collected over a wide geographic area, including the Baja California peninsula, and mainland localities in southern Arizona, USA and Sonora, Mexico, showed several well-supported partitions corresponding to Cx. quinquefasciatus, Cx. tarsalis, and two unidentified species, Culex sp. 1 and sp. 2. Culex quinquefasciatus was found at all localities and was the most abundant species collected. Culex tarsalis was collected only at Tucson, Arizona and Guaymas, Sonora. The two unidentified species of Culex were most abundant at Navojoa in southern Sonora. Haplotype and nucleotide diversities in the COI gene segment were substantially lower in Cx. quinquefasciatus compared with the other three species. Analysis of molecular variance revealed little structure among seven populations of Cx. quinquefasciatus, whereas significant structure was found between the two populations of Cx. tarsalis. Evidence for an historical population expansion beginning in the Pleistocene was found for Cx. tarsalis. Possible explanations for the large differences in genetic diversity between Cx. quinquefasciatus and the other species of Culex are presented. PMID:24302868

Objective To determine the mosquito repellent activity of herbal essential oils against female Aedes aegypti and Culex quinquefasciatus. Methods On a volunteer's forearm, 0.1 mL of each essential oil was applied to 3 cm×10 cm of exposed skin. The protection time was recorded for 3 min after every 30 min. Results Essential oil from clove oil in olive oil and in coconut oil gave the longest lasting period of 76.50 min and 96.00 min respectively against Aedes aegypti. The citronella grass oil in olive oil, citronella grass oil in coconut oil and lemongrass oil in coconut oil exhibited protection against Culex quinquefasciatus at 165.00, 105.00, and 112.50 min respectively. Conclusions The results clearly indicated that clove, citronella and lemongrass oil were the most promising for repellency against mosquito species. These oils could be used to develop a new formulation to control mosquitoes.

Three participants with callosal agenesis and 12 neurologically normal participants were tested on a simple reaction time task, with visual evoked potentials collected using a high-density 128-channel system. Independent-components analyses were performed on the averaged visual evoked potentials to isolate the components of interest. Contrary to previous research with acallosals, evidence of ipsilateral activation was present in all 3 participants. Although ipsilateral visual components were present in all 4 unilateral conditions in the 2 related acallosal participants, in the 3rd, these were present only in the crossed visual field-hand conditions and not in the uncrossed conditions. Suggestions are made as to why these results differ from earlier findings and as to the neural mechanisms facilitating this ipsilateral activation.

strains of Aedes albopictus. Ae. aegypti, and Culex tritaeniorhynchus. Female mosquitoes were infected by intrathoracic inoculation with WN virus , and the...of Cx. tritae- periment. West Nile virus infection of the pa- niorhYnchus.Ae. albopcwtus, and Ae. aeg’ypi was rental female mosquitoes in all... females in experiment no. i we e infected with stock virus that had received 16 intracerebral passailes in mice and four intsthoeiaeie pauffiie in Cx

Resistant strains of Culex quinquefasciatus were selected in the larval stage by exposure to sublethal doses of DDT. The size, body weight and fecundity of DDT-resistant mosquitoes were significantly decreased. There was no significant difference between the longevity of females in the original strain and the DDT selected strains, however, there was a significant difference in the males of the DDT-resistant strains than those of the original strain.

Motion visual evoked potentials (motion VEPs) have been used since the late 1960s to investigate the properties of human visual motion processing, and continue to be a popular tool with a possible future in clinical diagnosis. This review first provides a synopsis of the characteristics of motion VEPs and then summarizes important methodological aspects. A subsequent overview illustrates how motion VEPs have been applied to study basic functions of human motion processing and shows perspectives for their use as a diagnostic tool.

The local abundance of Culex mosquitoes is a central factor adding to the risk of West Nile virus transmission, and vector abundance data influence public health decisions. This study evaluated differences in abundance estimates from mosquitoes trapped using two common methods: CO2-baited CDC light traps and infusion-baited gravid traps in suburban, Chicago, Illinois. On a weekly basis, the two methods were modestly correlated (r = 0.219) across 71 weeks over 4 years. Lagged weather conditions of up to four weeks were associated with the number of mosquitoes collected in light and gravid traps. Collections in light traps were higher with higher temperature in the same week, higher precipitation one, two, and four weeks before the week of trapping, and lower maximum average wind speed. Collections in gravid traps were higher with higher temperature in the same week and one week earlier, lower temperature four weeks earlier, and with higher precipitation two and four weeks earlier. Culex abundance estimates from light traps were significantly higher in semi-natural areas compared to residential areas, but abundance estimates from gravid traps did not vary by the landscape type. These results highlight the importance of the surveillance methods used in the assessment of local Culex abundance estimates. Measures of risk of exposure to West Nile virus should assess carefully how mosquito abundance has been estimated and integrated into assessments of transmission risk. PMID:27375359

Introduction Amplification and transmission of West Nile virus (WNV) by mosquitoes are driven by presence and number of viraemic/susceptible avian hosts. Methods In order to predict risk of WNV infection to humans, we collected mosquitoes from horse stables in Lagos and Ibadan, southwestern Nigeria. The mosquitoes were sorted and tested in pools with real-time RT-PCR to detect WNV (or flavivirus) RNA using WNV-specific primers and probes, as well as, pan-flavivirus-specific primers in two-step real-time RT-PCR. Minimum infection rate (MIR) was used to estimate mosquito infection rate. Results Only two genera of mosquitoes were caught (Culex, 98.9% and Aedes, 1.0%) totalling 4,112 females. None of the 424 mosquito pools tested was positive for WNV RNA; consequently the MIR was zero. Sequencing and BLAST analysis of amplicons detected in pan-flavivirus primer-mediated RT-PCR gave a consensus sequence of 28S rRNA of Culex quinquefasciatus suggesting integration of flaviviral RNA into mosquito genome. Conclusion While the latter finding requires further investigation, we conclude there was little or no risk of human infection with WNV in the study areas during sampling. There was predominance of Culex mosquito, a competent WNV vector, around horse stables in the study areas. However, mosquito surveillance needs to continue for prompt detection of WNV activity in mosquitoes. PMID:27279943

Somatosensory evoked potential (SSEP), evoked spinal cord potential (ESCP) and motor evoked potential (MEP) have been used to detect spinal cord ischemia during aortic surgery. SSEP evaluates the sensory pathway, and is recorded from the sensory cortex by peripheral nerve stimulation. The interval from the onset of ischemia to change is relatively long(5-10 minutes). It has less frequently been used because of the high false negative and false positive rate. ESCP is recorded from the spinal cord by direct stimulation of the cord. It reflects the function of spinal tract but not that of alpha motor neurons. It is resistant to anesthesia and both the sensitivity and specificity is high, but the interval from ischemia to change is relatively long. Together with the necessity of 2 epidural electrodes, its application in aortic surgery has become infrequent. Since the introduction of train pulse transcranial electrical stimulation, myogenic MEP have gained widespread acceptance. It evaluates motor pathways from the cortex to the muscle, and therefore is influenced by non-spinal factors such as peripheral nerve ischemia. Its vulnerability to anesthesia requires special anesthetic consideration, and baseline amplitude fluctuation is common. It is highly sensitive and shows changes in the early phase of spinal cord ischemia.

Some species of mosquito can detect the presence of larvivorous fish and select against ovipositing in pools supporting them. The effect of kairomones released by the crimson-spotted rainbowfish Melanotaenia duboulayi (Castelnau) on the oviposition behavior of the freshwater mosquitoes Culex annulirostris Skuse, Culex quinquefasciatus Say, and Aedes notoscriptus Skuse was evaluated in both laboratory and outdoor artificial pool experiments. In the laboratory, colony-reared Cx. annulirostris selected against ovipositing in water that had contained M. duboulayi at densities of one fish per 5, 30, 180, and 1,080 liters. In contrast, gravid Ae. notoscriptus showed a preference for water that had contained the highest density of M. duboulayi (1 in 5 liters). Gravid Cx. quinquefasciatus were neither repelled nor attracted to water that had previously contained M. duboulayi. In outdoor artificial pool experiments, wild Cx. annulirostris females selected against ovipositing in pools containing caged M. duboulayi stocked at rates of 1, 4, and 10 g per 1,000 liters. When fish were removed from the pools, the repellent effect persisted for at least 24 h, whereas at the lower stocking rate, an avoidance response was not noted until 48 h after fish were placed in pools. This suggests a volatile substance, however, the exact nature of the kairomone/s has not been identified.

We assessed octylphenol (OP), an estrogenic endocrine-disrupting chemical, and UV-B radiation, a known stressor in amphibian development, for their effects on hypothalamic gene expression and premetamorphic development in the leopard frog Rana pipiens. Newly hatched tadpoles were exposed for 10 days to OP alone at two different dose levels; to subambient UV-B radiation alone; and to two combinations of OP and UV-B. Control animals were exposed to ethanol vehicle (0.01%) exposure, a subset of tadpoles from each treatment group was raised to metamorphosis to assess differences in body weight and time required for hindlimb emergence. Tadpoles from one of the OP/UV-B combination groups had greater body weight and earlier hindlimb emergence (p < 0.05), but neither OP nor UV-B alone produced significant changes in body weight or hindlimb emergence, indicating a potential mechanism of interaction between OP and UV-B. We hypothesized that the developing hypothalamus might be a potential environmental sensor for neurotoxicologic studies because of its role in the endocrine control of metamorphosis. We used a differential display strategy to identify candidate genes differentially expressed in the hypothalamic region of the exposed tadpoles. Homology cloning was performed to obtain R. pipiens glutamate decarboxylases--GAD65 and GAD67, enzymes involved in the synthesis of the neurotransmitter gamma-aminobutyric acid (GABA). cDNA expression profiles revealed that OP and UV-B affected the levels of several candidate transcripts in tadpole (i.e., Nck, Ash, and phospholipase C gamma-binding protein 4 and brain angiogenesis inhibitor-3) and metamorph (i.e., GAD67, cytochrome C oxidase, and brain angiogenesis inhibitor-2 and -3) brains. This study represents a novel approach in toxicology that combines physiologic and molecular end points and indicates that levels of OP commonly found in the environment and subambient levels of UV-B alter the expression of important hypothalamic

Laser-evoked potentials are widely used to investigate nociceptive pathways. The newly developed contact heat stimulator for evoking brain response has the advantages of obtaining reliable scalp potentials and absence of cutaneous lesions. This study aimed to identify the most appropriate stimulation site with consistent cortical responses, and to correlate several parameters of the contact heat evoked potentials (CHEPs) with age, gender, and body height in normal subjects. CHEPs were recorded at Cz with a contact heat stimulator (Medoc, Israel) in 35 normal controls. The subjects were asked to keep eyes open and remain alert. The baseline temperature was 32 degrees C, and stimulation peak heat intensity of 51 degrees C was applied to five body sites: bilateral forearm, right dorsum hand, right peroneal area, and right dorsum foot. Reproducible CHEPs were recorded more frequently when stimulated at volar forearm (62.5%) than at the lower limbs (around 40%). The first negative peak latency (N1) was 370.1 +/- 20.3 ms, first positive peak latency (P1) was 502.4 +/- 33.0 ms, and peak to peak amplitude was 10.2 +/- 4.9 microV with stimulation of the forearm. Perceived pain intensity was not correlated with the presence or amplitude of CHEPs. No gender or inter-side differences were observed for N1 latency and N1-P1 amplitude. Also, no correlation was noted between N1 and age or body height. These results support future clinical access of CHEPs as a diagnostic tool.

Fragrances, such as plant odors, have been shown to evoke autonomic response patterns associated with Ekman's (Ekman et al., 1983) basic emotions happiness, surprise, anger, fear, sadness, and disgust. Inducing positive emotions by odors in highly frequented public spaces could serve to improve the quality of life in urban environments. Thus, the present study evaluated the potency of ambient odors connoted with an urban environment to evoke basic emotions on an autonomic and cognitive response level. Synthetic mixtures representing the odors of disinfectant, candles/bees wax, summer air, burnt smell, vomit and musty smell as well as odorless water as a control were presented five times in random order to 30 healthy, non-smoking human subjects with intact sense of smell. Skin temperature, skin conductance, breathing rate, forearm muscle activity, blink rate, and heart rate were recorded simultaneously. Subjects rated the odors in terms of pleasantness, intensity and familiarity and gave verbal labels to each odor as well as cognitive associations with the basic emotions. The results showed that the amplitude of the skin conductance response (SCR) varied as a function of odor presentation. Burnt smell and vomit elicited significantly higher electrodermal responses than summer air. Also, a negative correlation was revealed between the amplitude of the SCR and hedonic odor valence indicating that the magnitude of the electrodermal response increased with odor unpleasantness. The analysis of the cognitive associations between odors and basic emotions showed that candles/bees wax and summer air were specifically associated with happiness whereas burnt smell and vomit were uniquely associated with disgust. Our findings suggest that city odors may evoke specific cognitive associations of basic emotions and that autonomic activity elicited by such odors is related to odor hedonics. PMID:24860522

Prior to acquisition of the first host blood meal, the anautogenous mosquito Culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. In the current study, we conducted whole transcriptome analyses of adult female Culex mosquitoes to iden...

Clicks and other acoustic stimuli evoke time-locked responses from the brain of man. The properties of the waves recordable within the interval from 1 to 10 msec after the stimuli strike the eardrum are discussed along with factors influencing the waves in the 100 to 500 msec epoch. So-called brainstem responses from a normal young adult are considered. No waves were observed for clicks to weak to be heard. With increasing stimulus strength the waves become larger in amplitude and their latency shortens.

Retrieval from semantic memory is usually considered within a time window around 300–600 ms. Here we suggest that lexical access already occurs at around 100 ms. This interpretation is based on the finding that semantically rich and frequent words exhibit a significantly shorter topographical latency difference between the site with the shortest P1 latency (leading site) and that with the longest P1 latency (trailing site). This latency difference can be described in terms of an evoked traveling alpha wave as was already shown in earlier studies. PMID:24486978

Relationship between evoked activity and spontaneous activity in neuronal circuits is one of the important theme for the improvement of neuroprosthetic apparatus. The spontaneous activity and evoked action potentials are mutually related in the cultured neuronal network autonomously reconstructed on the culture dish, but there is a question whether spontaneous activity and the evoked action potentials constitute one state respectively or the spontaneous activity is only a random background noise. Comparing the frequencies and standard deviations of spontaneous activity with those of evoked activity, we found that the silent and reproducible period lasting for 1 sec immediately after the activity evoked primally. In addition, the repetitive stimuli suppress the spontaneously occurring bursting activity in frequency, even though the inter-stimulus-interval was more than 10 sec. These results suggests that distinct internal state of the neuronal circuit was triggered by an electrical stimulation, and there were state of spontaneous mode and evoked mode in a neuronal circuit.

Observations on Culex quinquefasciatus Say in Samoa during a study of the epidemiology and control of subperiodic bancroftian filariasis are reported. The man-biting rate of C. quinquefasciatus was comparable with that of Aedes (Finlaya) samoanus Gruenberg in one indicator village and lower in another. The house-frequenting behaviour of the two species was similar. Culex quinquefasciatus was active throughout the night with peak biting during midnight hours, and its survivorship, estimated by two methods, was higher than those of the main vectors, A. (Stegomyia) polynesiensis Marks and A. samoanus. Culex quinquefasciatus showed low susceptibility to subperiodic Wuchereria bancrofti, compared with the two main vectors, in laboratory feeding experiments on volunteers with three levels of microfilaraemia. This agrees with the observed very low natural infection rate, suggesting that C. quinquefasciatus is an inefficient vector of subperiodic W. bancrofti in Samoa.

Wild-caught adult Rana pipiens females were captured in midsummer and fed diets of crickets, flies sowbugs or wax moth larvae during a three-month period of active vitellogenesis. The cricket diet supported the most extensive body weight gain during this time and promoted a prolonged period of weight increase in an additional long-term study. Synchronous growth of the oocytes occurred in all four groups, but the ovaries and oviducts of cricket-fed animals were significantly larger than those of frogs on the other three diets. The significantly higher liver weights of frogs fed wax moth larvae may have reflected an augmentation of hepatic energy stores. Fat body weights were also highest in this group of animals. Frogs fed crickets and wax moth larvae possessed larger fat bodies than did the midsummer control animals killed immediately after their arrival in the laboratory. In contrast, frogs fed flies and sowbugs had smaller fat bodies than did the initial controls, suggesting that animals on these diets had utilized fat body lipid during vitellogenesis. Gastrocnemius and final body weights were lowest in frogs fed wax moth larvae. These findings may have reflected the nutritional content of the diet or the reduction in appetite frequently noted in these animals during observations of feeding behavior.

In the past 30 years, many amphibian species have suffered population declines throughout the world. Mass mortality have been frequently reported, and in several instances, infectious diseases appear to be the cause of death. The role that contaminants could play in these die-offs through immunotoxic effects has been poorly investigated. In this study, juvenile leopard frogs (Rana pipiens) were exposed for 21 d to a mixture of six pesticides (atrazine, metribuzin, aldicarb, endosulfane, lindane, and dieldrin) and subsequently challenged with a parasitic nematode, Rhabdias ranae. Exposure to the mixture at environmentally realistic concentrations significantly reduced lymphocyte proliferation. Three weeks after the end of the exposure, lymphocyte proliferation had recovered and was stimulated in frogs challenged with parasites with the exception of those previously exposed to the highest concentration. No pesticide effects on phagocytosis and splenocyte numbers were detectable at the end of the exposure period, but these two parameters were diminished 21 d after the infection challenge in frogs previously exposed to the highest levels of pesticides. In these animals, the prevalence of lung infection by R. ranae also tended to be higher. These results suggest that agricultural pesticides can alter the immune response of frogs and affect their ability to deal with parasitic infection.

The external surface of the cornea and adjacent epidermis of larvae in representative developmental stages and of adult frogs, Rana pipiens, was studied by scanning electron microscopy. Surface cells are polygonal, usually hexagonal, in outline and covered with microprojections. During larval development prior to metamorphic stages, neither eyelids nor Harderian glands have developed; microprojections on the corneal surface are high and branched, and cell boundaries are elevated. On the anterior portion of the cornea and on the epidermis near the eye, the surface pattern is less dense, and ciliated cells are present. During metamorphic stages, corneal cell boundaries become less prominent and the pattern of microprojections more variable and markedly different from that of larvae of earlier stages. Corneal cells have a spongy appearance, are covered by a coating material, or are characterized as light or dark based on their brightness and surface texture. As eyelids develop in metamorphic stages XX-XXI, the numbers of ciliated cells increase dramatically, both on the corneal surface and on the edges of the developing lids. In later metamorphic stages XXII to XXV, lids and Harderian glands become well-developed, and cilia are no longer observed. The adjacent epidermal surface becomes devoid of cilia but perforated by openings of cutaneous glands. Its spongy appearance is similar to that of both the cornea and neighboring epidermis of the mature frog. Changes in corneal surface features are probably metamorphic events associated with development of lids and Harderian glands and a shift from an aqueous to an air environment.

Although increasing evidence links plana chlorinated hydrocarbons, such as polychlorinated biphenyls (PCBs), to decreases in survival and reproduction of fish, mammals, and birds near Green Bay, Wisconsin, and the Great Lakes, USA, relatively little is known of their bioaccumulation or of their possible effects in amphibians. The authors exposed embryos and larvae of two ranid species commonly occurring in the Green Bay ecosystem, the green frog (Rana clamitans) and the leopard frog (Rana pipiens), to PCB 126, a model coplanar PCB compound. Nominal concentrations ranged from 0.005 to 50 {micro}g/L, and exposure lasted through metamorphosis. Tissue concentrations of PCB 126 in tadpoles that did not metamorphose by the end of the experiment ranged from 1.2 to 9,600 ng/g wet mass. No significant mortality of embryos occurred before hatching; however, survival of larvae was significantly reduced at the highest concentration for both species. Few deformities were observed, but the incidence of edema was significantly higher in tadpoles exposed to 50 {micro}g/L. Swimming speed and growth of tadpoles was also significantly reduced in this treatment. The percent of tadpoles that reached metamorphosis was significantly lower in green frogs at the highest concentration, and no leopard frogs survived past day 47 of the experiment in this treatment. At high concentrations, PCB 126 affected both ranid species; however, sublethal effects were not apparent for the parameters the authors measured at concentrations that occur in water in the Green Bay ecosystem.

Background Reports of malformed frogs have increased throughout the North American continent in recent years. Most of the observed malformations have involved the hind limbs. The goal of this study was to accurately characterize the hind limb malformations in wild frogs as an important step toward understanding the possible etiologies. Methods During 1997 and 1998, 182 recently metamorphosed northern leopard frogs (Rana pipiens) were collected from Minnesota, Vermont, and Maine. Malformed hind limbs were present in 157 (86%) of these frogs, which underwent necropsy and radiographic evaluation at the National Wildlife Health Center. These malformations are described in detail and classified into four major categories: (1) no limb (amelia); (2) multiple limbs or limb elements (polymelia, polydactyly, polyphalangy); (3) reduced limb segments or elements (phocomelia, ectromelia, ectrodactyly, and brachydactyly; and (4) distally complete but malformed limb (bone rotations, bridging, skin webbing, and micromelia). Results Amelia and reduced segments and/or elements were the most common finding. Frogs with bilateral hind limb malformations were not common, and in only eight of these 22 frogs were the malformations symmetrical. Malformations of a given type tended to occur in frogs collected from the same site, but the types of malformations varied widely among all three states, and between study sites within Minnesota. Conclusions Clustering of malformation type suggests that developmental events may produce a variety of phenotypes depending on the timing, sequence, and severity of the environmental insult. Hind limb malformations in free-living frogs transcend current mechanistic explanations of tetrapod limb development.

The present study aims to investigate the susceptibility status of Aedes albopictus, Culex quinquefasciatus, and Cx. vishnui collected from a pig farm in Tanjung Sepat, Selangor, toward 11 insecticides representing the classes of organochlorines, carbamates, organophosphates, and pyrethroids. The results of a World Health Organization adult mosquito bioassay revealed that Ae. albopictus, Cx. quinquefasciatus, and Cx. vishnui exhibited different susceptibility toward various insecticides. Overall, pyrethroids were able to induce rapid knockdown for all test mosquito species. The pyrethroids lambdacyhalothrin and etofenprox were able to cause high mortality (> 80%) of all 3 species. The findings of the present study will benefit local authorities in selecting appropriate dosage of insecticides to be used in mosquito control in this area.

Thoracoabdominal aortic disease (aneurysm or dissection) has increased in recent decades. Surgery is the curative treatment but is associated to high perioperative morbidity and mortality risks. Paraplegia is one of the most severe complications, whose incidence has decreased significantly with the implementation of spinal cord protection strategies. No single method or combination of methods has proven to be fully effective in preventing paraplegia. This review is intended to analyse the scientific evidence available on the role of intraoperative monitoring with motor evoked potentials in the neurological outcome of patients undergoing thoracoabdominal aortic surgery. An online search (PubMed) was conducted. Relevant references were selected and reviewed. Intraoperative monitoring with motor evoked potentials (MEP) allows early detection of ischemic events and a targeted intervention to prevent the development of spinal cord injury, significantly reducing the incidence of postoperative paraplegia. MEP monitoring may undergo several intraoperative interferences which may compromise their interpretation. Neuromuscular blockade is the main limiting factor of anesthetic origin. It is essential to strike a balance between monitoring conditions and surgical and anesthetic needs as well as to evaluate the risks and benefits of the technique for each patient. MEP monitoring improves neurological outcome when integrated in a multidisciplinary strategy which must include multiple protective mechanisms that should be tailored to each hospital reality.

The auditory sensory pathway has been studied in meditators, using midlatency and short latency auditory evoked potentials. The present study evaluated long latency auditory evoked potentials (LLAEPs) during meditation. Sixty male participants, aged between 18 and 31 years (group mean±SD, 20.5±3.8 years), were assessed in 4 mental states based on descriptions in the traditional texts. They were (a) random thinking, (b) nonmeditative focusing, (c) meditative focusing, and (d) meditation. The order of the sessions was randomly assigned. The LLAEP components studied were P1 (40-60 ms), N1 (75-115 ms), P2 (120-180 ms), and N2 (180-280 ms). For each component, the peak amplitude and peak latency were measured from the prestimulus baseline. There was significant decrease in the peak latency of the P2 component during and after meditation (P

California statewide West Nile virus (WNV) minimum infection rates in Culex thriambus Dyar mosquitoes are high; however, few specimens are submitted and tested each year, as their distribution seems limited to larval habitats along riparian systems. To evaluate the role of Cx. thriambus in the amplification, maintenance, and overwintering of WNV in Lake County, CA, the bionomics and vector potential of the species was investigated during 2014 and 2015. Culex thriambus was the most abundant mosquito species, with 1,153 adults and 7,624 immatures collected by vacuum aspiration and dip sampling, respectively, at the primary study site. Detection of WNV in four mosquito pools during September through November coincided with peak seasonality. Females entered and maintained a reproductive diapause during winter under field and seminatural conditions. Diapause was initiated in the majority of Cx. thriambus females by October and was terminated by 30 March. Some parous females (7.1%) and those in host-seeking arrest (7.1%) were collected throughout the winter period. An accrual of 679.51 degree-days (°D) was necessary for diapause termination under seminatural conditions. Culex thriambus females fed on 16 different avian species during spring and summer, and no mammalian feeds were detected. West Nile viral RNA was detected in four of 42 Cx. thriambus pools tested during June through November and infection rates ranged from 3.53–28.15/1,000 tested. In summary, WNV transmission may be increased along riparian corridors throughout California where Cx. thriambus mosquitoes remain relatively abundant. PMID:27493251

The subgenus Melanoconion is the second largest subgenus within the genus Culex, with 160 described species. Several of the species are proven vectors of arboviruses, including West Nile virus, Venezuelan equine encephalitis virus complex and Eastern equine encephalomyelitis virus. Species of Melanoconion are well distributed from southern North America to most countries of South America and display the highest species diversity in tropical regions. Taxonomical identification within this group has been primarily based on morphological characters, with the male genitalia as the source of the most solid diagnostic features. The difficulty in reaching accurate species determinations when studying specimens of Culex (Melanoconion) has been extensively documented as a real limitation to expand knowledge of these insects. We tested the utility of the mitochondrial gene COI as a complementary tool in the taxonomy of Melanoconion. Using a data set of 120 COI sequences from Culex specimen captured in several localities in Brazil, the utility of COI barcodes for species delimitation is discussed through the evaluation of genetic divergences among specimens and the clustering patterns of species in three topologies obtained with Neighbor Joining, Maximum Likelihood and Bayesian phylogenetic inference. For all specimens included in this study a previous morphological examination was performed, and most of the taxonomical determinations were corroborated using the COI barcode. We generated COI sequences that belong to 48 species of Melanoconion, with a mean intraspecific K2P genetic divergence of 3%; and all interspecific divergence values higher than the intraspecific divergence values. This is the first comprehensive study of subgenus Melanoconion, with evidence of COI as a useful and accessible DNA barcode.

A strategy to engineer a strain of Culex mosquitoes refractory to filarial transmission is described. A requirement for success of the strategy is identification of a flight muscle-specific promoter that functions in the mosquito. A GFP marker gene under the control of the promoter region of the Drosophila melanogaster act88F gene was inserted into the genome of Culex quinquefasciatus. Transformation was confirmed by Mendelian genetics. Hybridization of a genomic Southern blot to a radiolabeled probe verified that the entire donor plasmid integrated into the mosquito genome. GFP expression in the transgenic mosquitoes was restricted to the flight muscles.

To control West Nile virus (WNV), it is necessary to know which mosquitoes are able to transmit this virus. Therefore, we evaluated the WNV vector potential of several North American mosquito species. Culex restuans and Cx. salinarius, two species from which WNV was isolated in New York in 2000, were efficient laboratory vectors. Cx. quinquefasciatus and Cx. nigripalpus from Florida were competent but only moderately efficient vectors. Coquillettidia perturbans was an inefficient laboratory vector. As WNV extends its range, exposure of additional mosquito species may alter its epidemiology. PMID:11747732

The mosquito larvicide methoprene is a juvenile growth hormone mimic that is widely used to control mosquitoes. This chemical disrupts normal mosquito development, drastically inhibiting emergence from the pupal to the adult stage. If the presence of methoprene attracts or deters mosquitoes from ovipositing it could have implications for mosquito control. This study evaluates whether methoprene attracts or deters mosquitoes likely to oviposit in catch basins. In a field experiment, methoprene formulated as liquid larvicide did not affect oviposition of either Culex spp. or Aedes japonicus in 19 liter plastic buckets.

Why do we listen to sad music? We seek to answer this question using a psychological approach. It is possible to distinguish perceived emotions from those that are experienced. Therefore, we hypothesized that, although sad music is perceived as sad, listeners actually feel (experience) pleasant emotions concurrent with sadness. This hypothesis was supported, which led us to question whether sadness in the context of art is truly an unpleasant emotion. While experiencing sadness may be unpleasant, it may also be somewhat pleasant when experienced in the context of art, for example, when listening to sad music. We consider musically evoked emotion vicarious, as we are not threatened when we experience it, in the way that we can be during the course of experiencing emotion in daily life. When we listen to sad music, we experience vicarious sadness. In this review, we propose two sides to sadness by suggesting vicarious emotion. PMID:24910621

The techniques of cognitive evoked potentials are considered long and technically complex, which is why their use in clinical practice is not very widespread in spite of their potential utility. Recent advances in registering and analysis, together with improvement of the software managing these signals, have appreciably reduced these problems. Mismatch negativity stands out as the most promising of all the cognitive potentials due to its special characteristics regarding its generation requisites and its proven clinical utility. The fact that it can be generated without care requirements makes it especially useful for evaluating subjects with a low level of consciousness; it serves for predicting when they will emerge from a coma, amongst other uses. The incorporation of this technique into the arsenal of neurophysiological techniques for evaluating the state of these subjects will bring a substantial improvement in the evaluation of cases whose management in clinical practice is extremely complex.

Night vision goggles (NVG's) have widespread use in military and civilian environments. NVG's amplify ambient illumination making performance possible when there is insufficient illumination for normal vision. While visual performance through NVG's is commonly assessed by measuring threshold functions such as visual acuity, few attempts have been made to assess vision through NVG's at suprathreshold levels of stimulation. Such information would be useful to better understand vision through NVG's across a range of stimulus conditions. In this study visual evoked potentials (VEP's) were used to evaluate vision through NVG's across a range of stimulus contrasts. The amplitude and latency of the VEP varied linearly with log contrast. A comparison of VEP's recorded with and without NVG's was used to estimate contrast attenuation through the device. VEP's offer an objective, electrophysiological tool to assess visual performance through NVG's at both threshold and suprathreshold levels of visual stimulation.

The objective of this study was to evaluate the association between rest heart rate (HR) and the components of the auditory evoked-related potentials (ERPs) at rest in women. We investigated 21 healthy female university students between 18 and 24 years old. We performed complete audiological evaluation and measurement of heart rate for 10 minutes at rest (heart rate monitor Polar RS800CX) and performed ERPs analysis (discrepancy in frequency and duration). There was a moderate negative correlation of the N1 and P3a with rest HR and a strong positive correlation of the P2 and N2 components with rest HR. Larger components of the ERP are associated with higher rest HR. PMID:26504838

Two evoked potential (EP) techniques and the selective reminding test were employed to investigate an apparently benign forgetfulness in seven elderly subjects and seven age-matched elderly subjects with normal memory. EPs were also recorded in a group of seven young adults. Latency of the P3 component, which has been demonstrated to increase in primary degenerative dementia, displayed the normal age-related variation in both elderly groups, but did not differ between the forgetful subjects and the elderly controls. Further, no difference in the recovery cycle of the EP, as measured in a two tone stimulation paradigm, was present between forgetful and elderly control groups. Reexamination of memory after nearly a year disclosed no evidence of deterioration in either elderly group. These findings suggest that senescent forgetfulness, as defined herein, may be a nonprogressive memory disorder.

We have developed a method for analyzing neural electromagnetic data that allows probabilistic inferences to be drawn about regions of activation. The method involves the generation of a large number of possible solutions which both fit the data and prior expectations about the nature of probable solutions made explicit by a Bayesian formalism. In addition, we have introduced a model for the current distributions that produce MEG (and EEG) data that allows extended regions of activity, and can easily incorporate prior information such as anatomical constraints from MRI. To evaluate the feasibility and utility of the Bayesian approach with actual data, we analyzed MEG data from a visual evoked response experiment. We compared Bayesian analyses of MEG responses to visual stimuli in the left and right visual fields, in order to examine the sensitivity of the method to detect known features of human visual cortex organization. We also examined the changing pattern of cortical activation as a function of time.

The authors developed a method for analyzing neural electromagnetic data that allows probabilistic inferences to be drawn about regions of activation. The method involves the generation of a large number of possible solutions which both fir the data and prior expectations about the nature of probable solutions made explicit by a Bayesian formalism. In addition, they have introduced a model for the current distributions that produce MEG and (EEG) data that allows extended regions of activity, and can easily incorporate prior information such as anatomical constraints from MRI. To evaluate the feasibility and utility of the Bayesian approach with actual data, they analyzed MEG data from a visual evoked response experiment. They compared Bayesian analyses of MEG responses to visual stimuli in the left and right visual fields, in order to examine the sensitivity of the method to detect known features of human visual cortex organization. They also examined the changing pattern of cortical activation as a function of time.

Why do we listen to sad music? We seek to answer this question using a psychological approach. It is possible to distinguish perceived emotions from those that are experienced. Therefore, we hypothesized that, although sad music is perceived as sad, listeners actually feel (experience) pleasant emotions concurrent with sadness. This hypothesis was supported, which led us to question whether sadness in the context of art is truly an unpleasant emotion. While experiencing sadness may be unpleasant, it may also be somewhat pleasant when experienced in the context of art, for example, when listening to sad music. We consider musically evoked emotion vicarious, as we are not threatened when we experience it, in the way that we can be during the course of experiencing emotion in daily life. When we listen to sad music, we experience vicarious sadness. In this review, we propose two sides to sadness by suggesting vicarious emotion.

Pattern reversal visual evoked potentials (pVEP) were studied in 39 male rubber factory workers in the age range of 18-55 years and 20 control subjects (aged 18-46 years) not exposed to the rubber factory environment. Results revealed that 20 (51%) rubber factory workers had abnormal latencies of wave P1 (dominant component of pVEP) as per accepted criteria of 99% tolerance limit set for the control group (i.e. any value above mean +3 SD of control was considered abnormal). The section-wise per cent distribution of abnormalities was vulcanization (83%), tubing (75%), calendering (60%), loading (38%) and mixing (14%). This study provides electrophysiological evidence that rubber factory environments affect the conduction processes in optical pathways from their origin in the retina to striate cortex. However, this study has its limitations in not identifying the specific chemical(s) causing these changes in VEP.

Intensive row crop agriculture (IRCA) for corn and soybean production is predominant in eastern and central North America. IRCA relies heavily on pesticide and nutrient inputs to maximize production under conventional systems. In 2003-2005, we assessed the occurrence of a suite of potential endocrine effects in amphibians inhabiting farm ponds and agricultural drains in IRCA areas of southwestern Ontario. Effects were compared to amphibians from two agricultural reference sites as well as four non-agricultural reference sites. Pesticide and nutrient concentrations were also determined in water samples from those sites. Atrazine and metolachlor were detected in most samples, exceeding 1 microg L(-1) at some sites. Blood samples were taken from northern leopard frogs (Rana pipiens) and green frogs (Rana clamitans) for analysis of circulating sex steroids and vitellogenin-like protein (Vtg-lp), a biomarker of exposure to environmental estrogens. Gonads were histologically examined for evidence of abnormalities. Some evidence of exposure to endocrine disrupting compounds was apparent from the data. The occurrence of testicular ovarian follicles (TOFS) in male R. pipiens was significantly higher (42%; p<0.05) at agricultural sites, particularly those in Chatham county compared to frogs from reference sites (7%). There was no difference in circulating sex steroid levels between frogs from agricultural and reference sites and sex steroid levels did not correlate with pesticide concentrations in the environment. No differences were detected in the gonadosomatic indices or stage of spermatogenesis between frogs from agricultural and non-agricultural regions (p>0.05). Plasma Vtg-lp was detected in only one male R. pipiens from an agricultural site. Neither gonad size, gonad maturity nor sex steroid levels differed between normal males and those with testicular oocytes. Although the proportion of testicular oocytes did not correlate directly with atrazine concentrations, it

In the experiments reported here, the amplitude and the latency of human compound action potentials (CAPs) evoked from a chirp stimulus are compared to those evoked from a traditional click stimulus. The chirp stimulus was created with a frequency sweep to compensate for basilar membrane traveling wave delay using the O-Chirp equations from Fobel and Dau [(2004). J. Acoust. Soc. Am. 116, 2213-2222] derived from otoacoustic emission data. Human cochlear traveling wave delay estimates were obtained from derived compound band action potentials provided by Eggermont [(1979). J. Acoust. Soc. Am. 65, 463-470]. CAPs were recorded from an electrode placed on the tympanic membrane (TM), and the acoustic signals were monitored with a probe tube microphone attached to the TM electrode. Results showed that the amplitude and latency of chirp-evoked N1 of the CAP differed from click-evoked CAPs in several regards. For the chirp-evoked CAP, the N1 amplitude was significantly larger than the click-evoked N1s. The latency-intensity function was significantly shallower for chirp-evoked CAPs as compared to click-evoked CAPs. This suggests that auditory nerve fibers respond with more unison to a chirp stimulus than to a click stimulus.

Gravid mosquito traps are commonly used for both arbovirus surveillance and population surveillance of mosquitoes of the genus Culex. Oviposition substrates, used as baits in these traps, were tested against Culex under laboratory and field conditions. In the laboratory all substrates tested as 1% and 10% dilutions in 2-choice bioassays against female Cx. quinquefasciatus were significantly more effective than well water controls in eliciting oviposition. Strongest responses were to dilutions of dairy effluent, followed by larval water and infusions of alfalfa hay, alfalfa pellets, Bermuda hay, oak leaves, and Typha leaves, with lowest responses to cow manure infusion. In the field, few significant differences in collections were obtained between traps baited with different infusions. Significantly more Cx. quinquefasciatus and Cx. nigripalpus were collected in traps baited with cow manure infusion (highest) compared to alfalfa hay infusion (lowest). Responses of Cx. quinquefasciatus to dairy effluent and infusions of Bermuda hay, oak leaves, and Typha leaves were not significantly different from either cow manure infusion or alfalfa hay infusion. Responses of Cx. nigripalpus were highest to cow manure infusion and equally low to infusions of alfalfa hay and Typha leaves; moderate responses were observed to dairy effluent and infusions of Bermuda hay and oak leaves. Gravid females comprised 66.7-81.9% of the collections for each infusion type, with no significant difference among infusions in the proportion of gravid females collected.

Mosquito control is facing a threat because of the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. The purpose of the present study was to assess the effect of leaf ethyl acetate, acetone and methanol extracts of Aegle marmelos (L.) Correa ex Roxb (Rutaceae), Andrographis lineata Wallich ex Nees. (Acanthaceae), Andrographis paniculata (Burm.f.) Wall. ex Nees. (Acanthaceae), Cocculus hirsutus (L.) Diels (Menispermaceae), Eclipta prostrata L. (Asteraceae) and Tagetes erecta L. (Compositae) on ovicidal and oviposition-deterrent activities against Culex tritaeniorhynchus Giles (Diptera: Culicidae). The percentage of egg hatching in methanol extracts of Andrographis lineata, Cocculus hirsutus and T. erecta were 16, 12 and 16 exerted at 500 ppm, respectively. The percentage of effective oviposition repellency was 97.77 at 500 ppm and the lowest repellency was 42.06 at 31.25 ppm in methanol and acetone extracts of Andrographis lineata and Andrographis paniculata, respectively. The oviposition activity index values revealed that the solvent plant extracts have deterrent effect, and they caused a remarkable negative response resulting in oviposition of very few eggs. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal ecofriendly approach for the control of the Culex tritaeniorhynchus.

Although Anopheles mosquitoes are the vectors for human Plasmodium spp., there are also other mosquito species-among them culicines (Culex spp., Aedes spp.)-present in malaria-endemic areas. Culicine mosquitoes transmit arboviruses and filarial worms to humans and are vectors for avian Plasmodium spp., but have never been observed to transmit human Plasmodium spp. When ingested by a culicine mosquito, parasites could either face an environment that does not allow development due to biologic incompatibility or be actively killed by the mosquito's immune system. In the latter case, the molecular mechanism of killing must be sufficiently powerful that Plasmodium is not able to overcome it. To investigate how human malaria parasites develop in culicine mosquitoes, we infected Culex quinquefasciatus with Plasmodium falciparum NF54 and monitored development of parasites in the blood bolus and midgut epithelium at different time points. Our results reveal that ookinetes develop in the midgut lumen of C. quinquefasciatus in slightly lower numbers than in Anopheles gambiae G3. After 30 hours, parasites have invaded the midgut and can be observed on the basal side of the midgut epithelium by confocal and transmission electron microscopy. Very few of the parasites in C. quinquefasciatus are alive, most of them are lysed. Eight days after the mosquito's blood meal, no oocysts can be found in C. quinquefasciatus. Our results suggest that the mosquito immune system could be involved in parasite killing early in development after ookinetes have crossed the midgut epithelium and come in contact with the mosquito hemolymph.

Culex mosquitoes have emerged as important model organisms for mosquito biology, and are disease vectors for multiple mosquito-borne pathogens, including West Nile virus. We characterized epoxide hydrolase activities in the mosquito Culex quinquefasciatus, which suggested multiple forms of epoxide hydrolases were present. We found EH activities on epoxy eicosatrienoic acids (EETs). EETs and other eicosanoids are well-established lipid signaling molecules in vertebrates. We showed EETs can be synthesized in vitro from arachidonic acids by mosquito lysate, and EETs were also detected in vivo both in larvae and adult mosquitoes by LC-MS/MS. The EH activities on EETs can be induced by blood feeding, and the highest activity was observed in the midgut of female mosquitoes. The enzyme activities on EETs can be inhibited by urea-based inhibitors designed for mammalian soluble epoxide hydrolases (sEH). The sEH inhibitors have been shown to play diverse biological roles in mammalian systems, and they can be useful tools to study the function of EETs in mosquitoes. Besides juvenile hormone metabolism and detoxification, insect epoxide hydrolases may also play a role in regulating lipid signaling molecules, such as EETs and other epoxy fatty acids, synthesized in vivo or obtained from blood feeding by female mosquitoes. PMID:25686802

Culex (Melanoconion) iolambdis (Dyar, 1918) is a mosquito (Diptera: Culicidae) species found throughout much of tropical America, including southern Florida. Relatively few reports are available regarding the ecology of Cx. iolambdis, despite its widespread distribution and putative involvement in transmission of Venezuelan equine encephalitis virus. To quantify habitat and host utilization, adults of Cx. iolambdis were sampled from resting shelters at a field site in Vero Beach, Florida, over a 12-month period. Culex iolambdis (1109 males, 3072 females) constituted more than half (56.76%) of all mosquitoes sampled (24 species) and was active year-round. Unfed females and gravid females of Cx. iolambdis were significantly more abundant in mangrove habitat, while males and blood-fed females were not. PCR-based bloodmeal analysis of 305 females revealed that Cx. iolambdis has very wide host breadth, feeding on birds (37.0% overall), reptiles (26.6%), amphibians (23.3%) and mammals (13.1%). Green heron (Butorides virescens), Southern leopard frog (Lithobates sphenocephala) and American alligator (Alligator mississippiensis) were the most commonly fed upon hosts. Bloodmeals from different host classes varied significantly with season, suggesting that Cx. iolambdis may play a role in the amplification and epidemic transmission of zoonotic arboviruses affecting human health.

About 1 million people in the world die each year from diseases spread by mosquitoes, and understanding the mechanism of host identification by the mosquitoes through olfaction is at stake. The role of odorant binding proteins (OBPs) in the primary molecular events of olfaction in mosquitoes is becoming an important focus of biological research in this area. Here, we present a comprehensive comparative genomics study of OBPs in the three disease-transmitting mosquito species Anopheles gambiae, Aedes aegypti, and Culex quinquefasciatus starting with the identification of 110 new OBPs in these three genomes. We have characterized their genomic distribution and orthologous and phylogenetic relationships. The diversity and expansion observed with respect to the Aedes and Culex genomes suggests that the OBP gene family acquired functional diversity concurrently with functional constraints posed on these two species. Sequences with unique features have been characterized such as the “two-domain OBPs” (previously known as Atypical OBPs) and “MinusC OBPs” in mosquito genomes. The extensive comparative genomics featured in this work hence provides useful primary insights into the role of OBPs in the molecular adaptations of mosquito olfactory system and could provide more clues for the identification of potential targets for insect repellants and attractants. PMID:23292137

Among members of the order Mononegavirales, RNA splicing events have been found only in the family Bornaviridae. Here, we report that a new rhabdovirus isolated from the mosquito Culex tritaeniorhynchus replicates in the nuclei of infected cells and requires RNA splicing for viral mRNA maturation. The virus, designated Culex tritaeniorhynchus rhabdovirus (CTRV), shares a similar genome organization with other rhabdoviruses, except for the presence of a putative intron in the coding region for the L protein. Molecular phylogenetic studies indicated that CTRV belongs to the family Rhabdoviridae, but it is yet to be assigned a genus. Electron microscopic analysis revealed that the CTRV virion is extremely elongated, unlike virions of rhabdoviruses, which are generally bullet shaped. Northern hybridization confirmed that a large transcript (approximately 6,500 nucleotides [nt]) from the CTRV L gene was present in the infected cells. Strand-specific reverse transcription-PCR (RT-PCR) analyses identified the intron-exon boundaries and the 76-nt intron sequence, which contains the typical motif for eukaryotic spliceosomal intron-splice donor/acceptor sites (GU-AG), a predicted branch point, and a polypyrimidine tract. In situ hybridization exhibited that viral RNAs are primarily localized in the nucleus of infected cells, indicating that CTRV replicates in the nucleus and is allowed to utilize the host's nuclear splicing machinery. This is the first report of RNA splicing among the members of the family Rhabdoviridae. PMID:21507977

Visual evoked potentials (VEPs) are of great concern in cognitive and clinical neuroscience as well as in the recent research field of brain-computer interfaces (BCIs). In this study, a chirp-modulated stimulation was employed to serve as a novel type of visual stimulus. Based on our empirical study, the chirp stimuli visual evoked potential (Chirp-VEP) preserved frequency features of the chirp stimulus analogous to the steady state evoked potential (SSVEP), and therefore it can be regarded as a generalization of SSVEP. Specifically, we first investigated the characteristics of the Chirp-VEP in the time-frequency domain and the fractional domain via fractional Fourier transform. We also proposed a group delay technique to derive the apparent latency from Chirp-VEP. Results on EEG data showed that our approach outperformed the traditional SSVEP-based method in efficiency and ease of apparent latency estimation. For the recruited six subjects, the average apparent latencies ranged from 100 to 130 ms. Finally, we implemented a BCI system with six targets to validate the feasibility of Chirp-VEP as a potential candidate in the field of BCIs.

Visual evoked potentials (VEPs) are of great concern in cognitive and clinical neuroscience as well as in the recent research field of brain-computer interfaces (BCIs). In this study, a chirp-modulated stimulation was employed to serve as a novel type of visual stimulus. Based on our empirical study, the chirp stimuli visual evoked potential (Chirp-VEP) preserved frequency features of the chirp stimulus analogous to the steady state evoked potential (SSVEP), and therefore it can be regarded as a generalization of SSVEP. Specifically, we first investigated the characteristics of the Chirp-VEP in the time-frequency domain and the fractional domain via fractional Fourier transform. We also proposed a group delay technique to derive the apparent latency from Chirp-VEP. Results on EEG data showed that our approach outperformed the traditional SSVEP-based method in efficiency and ease of apparent latency estimation. For the recruited six subjects, the average apparent latencies ranged from 100 to 130 ms. Finally, we implemented a BCI system with six targets to validate the feasibility of Chirp-VEP as a potential candidate in the field of BCIs.

Anthropogenic introduction of species is a major contributor to loss of biodiversity. Translocations within the range of a species are less frequently recognized, but have the potential for negative effects as well. Genetic mixing may lead to loss of local adaptations or further decline through outbreeding depression. These cryptic invasions may be quite difficult to recognize, but genetic tools can be used to recognize and monitor such intraspecific introductions. Conversely, translocations within species can be an important conservation tool to reduce inbreeding depression and replace lost genetic diversity. Thus, cryptic invasions can be either an aid or a hindrance to conservation efforts. We tested for the presence of non-native genotypes and assessed the extent and nature of introgression in populations of Northern Leopard Frog (Rana pipiens) in the southwestern US, where populations have declined to a few remnant populations. The most abundant and diverse complex of populations in the region contained a mitochondrial haplotype that was not native to the western US, probably resulting from the introduction of released pets, laboratory animals, or release during fish stocking. These non-native haplotypes were well integrated into a large complex of ponds and lakes, contributing to high genetic diversity in this area. Logistically, the geographic extent of non-native genetic influence within this population precludes eliminating or controlling the non-native component of this population. We recommend assessing the progress and fate of the introgression over time—along with population fitness parameters—to determine whether this introduction is beneficial or detrimental to population persistence. Meanwhile, translocations from nearby locations with similar environmental conditions have the best prospects for avoiding problems with outbreeding depression in other declining populations and will also most effectively preserve regional genetic diversity.

For vectorborne infections, host selection by bloodfeeding arthropods dictates the interaction between host and pathogen. Because Culex mosquitoes that transmit West Nile virus (WNV) feed both on mammalian and avian hosts with varying competence, understanding the bloodfeeding patterns of these mosquitoes is important for understanding the transmission dynamics of WNV. Herein, we describe a new microsphere-based assay using Luminex xMAP® technology to rapidly identify 15 common hosts of Culex mosquitoes at our California study sites. The assay was verified with over 100 known vertebrate species samples and was used in conjunction with DNA sequencing to identify over 125 avian and mammalian host species from unknown Culex bloodmeals, more quickly and with less expense than sequencing alone. In addition, with multiplexed labeled probes, this microsphere array identified mixed bloodmeals that were difficult to discern with traditional sequencing. The microsphere set was easily expanded or reduced according to host range in a specific area, and this assay has made it possible to rapidly screen thousands of Culex spp. bloodmeals to extend our understanding of WNV transmission patterns. PMID:22136215

The aim of this study was to assess the occurrence of baculovirus infections in mosquitoes and characterize them by using molecular tools. Fortnightly collections were made of mosquito larvae in the city of Caraguatatuba. Six larvae of Culex quinquefasciatus were isolated that had white cysts (nodul...

CuniNPV is a member of the Dipteran–specific baculoviruses in the genus Deltabaculovirus that specifically infects mosquito larvae within the genus Culex while species of Aedes and Anopheles are refractory. Infections are restricted to the nuclei of larval midgut epithelial cells with transmission...

The Larvasonic™ Field Arm Mobile Wetlands Unit and SD-Mini were tested for efficacy against Culex larvae, and effects on aquatic nontarget organisms (NTO). The Field Arm provided 84.61% to 100% control of caged Culex larvae out to 0.91-m distance in shallow ditches and 60.45% control of Culex larvae at 0.61-m without any effects to caged NTO. Slow ditch treatment achieved 77.35% control compared to fast treatment (20.42%), whereas 77.65% control was obtained along edges of a neglected swimming pool, compared to near the middle (23.97%). In bucket tests, the SD-Mini provided >97% control of Culex and 85.35% reduction of immature giant water bugs, which decreased slightly (83.45%) over the monitoring period, which was not significantly different from cannibalistic damselflies (62.80%), with reduction of both being significantly higher than other NTO tested. There was a small (0.37%) reduction of dragonflies (naiads), due to cannibalism. Both Larvasonic units could effectively augment conventional larvicide operations in smaller areas without causing resistance within mosquito populations or harming NTO when used properly.

Sounds provide the most suitable stimuli for studies of information processes occurring in the brain during falling asleep and at different stages of sleep. The widely used analysis of evoked potentials averaged for groups of subjects has a number of disadvantages associated with their individual variability. Thus, in the present study, measures of the individual components of auditory evoked potentials were determined and selectively summed for individual subjects, with subsequent analysis by group. The aim of the present work was to identify measures of auditory evoked potentials providing quantitative assessment of the dynamics of the brain's functional state during the appearance of errors in activity associated with decreases in the level of waking and falling asleep. A monotonous psychomotor test was performed in the lying position with the eyes closed; this consisted of two alternating parts: the first was counting auditory stimuli from 1 to 10 with simultaneous pressing of a button, and the second was counting stimuli from 1 to 5 silently without pressing the button, and so on. Computer-generated sound stimuli (duration 50 msec, envelope filling frequency 1000 Hz, intensity 60 dB) were presented binaurally with interstimulus intervals of 2.4-2.7 sec. A total of 41 subjects took part (both genders, mean age 25 years), of which only 23 fell asleep; data for 14 subjects with sufficient episodes of falling asleep were analyzed. Comparison of measures of auditory evoked potentials (the latencies and amplitudes of the N1, P2, N2, and P3 components) during correct and erroneous psychomotor test trials showed that decreases in the level of consciousness elicited significant increases in the amplitudes of the components of the vertex N1-P2-N2 complex in series without button pressing. The greatest changes in auditory evoked potentials in both series were seen in the N2 component, with latency 330-360 msec, which has a common origin with the EEG theta rhythm and is

The visually evoked potential (VEP) was studied in normal and abnormal human subjects, and in Rhesus monkeys with central, paracentral, and peripheral photocoagulation lesions. A relatively simple protocol for clinical VEP testing is described. The monkeys showed similar VEP responses but these were smaller in amplitude than those obtained from human subjects. Central, but not paracentral or peripheral retinal lesions were associated with VEP abnormalities. For both monkey and human subjects, some variability of responses between normal and subjects was noted. Generally, there are differences in VEP responses obtained from the affected eye of abnormal subjects who had one eye which could serve as a control, as compared to responses from the normal eye. In these subjects as well as in subjects with two abnormal eyes, computer analysis of digitized VEP data from 10 Hz stimulus responses was performed. Fourier transformation analyses showed abnormalities which could be detected easily by evaluating the pattern of the amplitudes of the fundamental and first three harmonics. With this technique, it was possible to group correctly normal VEP's with eyes with normal visual acuity (greater than or equal to 20/30 or 0.67), and abnormal VEP's with eyes with poor visual acuity (less than 20/30 or 0.67) in 72% of cases. Analysis of the data obtained with 1 Hz and 10 Hz stimulation suggests that the components of the VEP related to visual acuity occur within the first 60-100 msec of the response, corresponding to the primary evoked response of Chiganek. The second, smaller wave of the response complex to 10 Hz flash stimuli corresponds to the primary evoked response, and is closely related to visual acuity. This was further supported in another series in which the digitized data was filtered around the stimulating frequency. It was possible to recognize visually this VEP waveform and subjectively interpret the record correctly in 85% of eyes with regard to visual acuity

The purpose of this review is to examine the validity of change in the cortical evoked response as a measure of inert gas narcosis in humans. Three criteria are defined which must all be met if a nonbehavioral measure is to be accepted as an indicator of narcosis. The evoked response is assessed in terms of these criteria. Two classes of experiments which have used the evoked response in hyperbaric ocnditions are identified. The first class allows the evoked response to be assessed against more than one of these criteria. The outcome of every experiment in this class supports the view that the evoked response is not a valid measure of narcosis. The second class of experiment assumed that the evoked response is a measure of narcosis and were not designed to assess validity appropriately. Arguments by Kinney and associates in support of the assumption of validity are shown to be unsound. Possible explanations for inability to demonstrate validity are discussed and it is suggested that factors other than narcotic potency of the breathing gas mixture determine or at least play a major role in determining amplitude of the evoked response.

Recent evidence suggests that infection by larvae of the trematode Ribeiroia ondatrae accounts for a significant proportion of limb malformations currently observed in amphibian populations of North America. However, the effects of R. ondatrae infection on northern leopard frogs (Rana pipiens), one of the species most frequently reported with malformations, have not been adequately explored. Moreover, the risk factors associated with R. ondatrae-induced malformations have not been clearly identified. We examined the effects of timing of infection on tadpole survival and limb development. Rana pipiens tadpoles were individually exposed to R. ondatrae cercariae at the pre-limb-bud (Gosner stages 24 and 25), limb-bud (Gosner stages 27 and 28), or paddle (Gosner stages 3133) stages of development and monitored through metamorphosis. The effects of infection were stage-specific. Infections acquired at the pre-limb-bud stage resulted in a high mortality rate (47.597.5%), whereas tadpoles infected at the limb-bud stage displayed a high malformation rate (16% overall), and the magnitude of effects increased with the level of exposure to cercariae. In contrast, infections acquired at the paddle stage had no effect on limb development or tadpole survival, which suggests that the timing of R. ondatrae infection in relation to the stage structure of tadpole populations in the wild is an important determinant of the degree to which populations are affected by R. ondatrae.

Auditory evoked potentials (AEP) were first used to examine hearing in elasmobranchs by Corwin and Bullock in the late 1970s and early 1980s, marking the first time AEPs had been measured in fishes. Results of these experiments identified the regions of the ear and brain in which sound is processed, though no actual hearing thresholds were measured. Those initial experiments provided the ground work for future AEP experiments to measure fish hearing abilities in a manner that is much faster and more convenient than classical conditioning. Data will be presented on recent experiments in which AEPs were used to measure the hearing thresholds of two species of elasmobranchs: the nurse shark, Ginglymostoma cirratum, and the yellow stingray, Urobatis jamaicencis. Audiograms were analyzed and compared to previously published audiograms obtained using classical conditioning with results indicating that hearing thresholds were similar for the two methods. These data suggest that AEP testing is a viable option when measuring hearing in elasmobranchs and can increase the speed in which future hearing measurements can be obtained.

Cough affects all individuals at different times, and its economic burden is substantial. Despite these widespread adverse effects, cough research relies on animal models, which hampers our understanding of the fundamental cause of cough. Postnasal drip is speculated to be one of the most frequent causes of chronic cough; however, this is a matter of debate. Here we show that mechanical stimuli by postnasal drip cause chronic cough. We distinguished human cough from sneezes and expiration reflexes by airflow patterns. Cough and sneeze exhibited one-peak and two-peak patterns, respectively, in expiratory airflow, which were also confirmed by animal models of cough and sneeze. Transgenic mice with ciliary dyskinesia coughed substantially and showed postnasal drip in the pharynx; furthermore, their cough was completely inhibited by nasal airway blockade of postnasal drip. We successfully reproduced cough observed in these mice by injecting artificial postnasal drip in wild-type mice. These results demonstrated that mechanical stimulation by postnasal drip evoked cough. The findings of our study can therefore be used to develop new antitussive drugs that prevent the root cause of cough. PMID:26581078

To evaluate the effect of stereotactic thalamotomy on the function of the corticospinal tract, we studied motor evoked potentials (MEPs) recorded by surface electromyography (EMG) in the left extensor carpi radialis (ECR) and flexor carpi radialis (FCR) with magnetic stimulation of the contralateral motor cortex in a 43-year-old patient with a severe postural and resting tremor of the left hand. The patient was diagnosed eight years previously with left hemiparkinsonism. The tremor was unresponsive to various medications. After thalamotomy the tremor had disappeared, confirmed by EMG studies. MEP latencies at rest were normal and did not change after thalamotomy. Volitional contraction of either ECR or FCR shortened the latency of the corresponding MEP before and after thalamotomy. However, before thalamotomy responses at rest were less well synchronized and followed by EMG silence with subsequent long duration tonic after discharges. Furthermore, during voluntary contraction the responses only slightly enhanced. After surgery MEPs at rest in both muscles were more synchronized and after-discharges had disappeared. Moreover, with volitional contraction of either ECR of FCR, the MEPs enhanced more dramatically. The silent periods (SPs) following the MEP during sustained voluntary contraction were longer after thalamotomy. The consistent MEP latencies suggest that the conduction of the pyramidal tract is unaffected by thalamotomy. The better synchronized responses, the alleviation of after-discharges and the longer SPs in this patient with hemiparkinsonism following thalamotomy suggest an improved sensorimotor integration, which may be the result of a reduced thalamic input onto suprasegmental levels.

Surface electrical stimulation (SES) is being investigated as a noninvasive method to evoke natural sensations distal to electrode location. This may improve treatment for phantom limb pain as well as provide an alternative method to deliver sensory feedback. The median and/or ulnar nerves of 35 subjects were stimulated at the elbow using surface electrodes. Strength-duration curves of hand sensation were found for each subject. All subjects experienced sensation in their hand, which was mostly described as a paresthesia-like sensation. The rheobase and chronaxie values were found to be lower for the median nerve than the ulnar nerve, with no significant difference between sexes. Repeated sessions with the same subject resulted in sufficient variability to suggest that recalculating the strength-duration curve for each electrode placement is necessary. Most of the recruitment curves in this study were generated with 28 to 36 data points. To quickly reproduce these curves with limited increase in error, we recommend 10 data points. Future studies will focus on obtaining different sensations using SES with the strength-duration curve defining the threshold of the effective parameter space.

Cough affects all individuals at different times, and its economic burden is substantial. Despite these widespread adverse effects, cough research relies on animal models, which hampers our understanding of the fundamental cause of cough. Postnasal drip is speculated to be one of the most frequent causes of chronic cough; however, this is a matter of debate. Here we show that mechanical stimuli by postnasal drip cause chronic cough. We distinguished human cough from sneezes and expiration reflexes by airflow patterns. Cough and sneeze exhibited one-peak and two-peak patterns, respectively, in expiratory airflow, which were also confirmed by animal models of cough and sneeze. Transgenic mice with ciliary dyskinesia coughed substantially and showed postnasal drip in the pharynx; furthermore, their cough was completely inhibited by nasal airway blockade of postnasal drip. We successfully reproduced cough observed in these mice by injecting artificial postnasal drip in wild-type mice. These results demonstrated that mechanical stimulation by postnasal drip evoked cough. The findings of our study can therefore be used to develop new antitussive drugs that prevent the root cause of cough.

The purpose of this study was to analyse the changes in transient evoked otoacoustic emissions (TEOAEs) with age. We studied 232 subjects above 60 years of age with a battery of audiological tests, including TEOAEs Our criterion for the presence of TEOAEs was based on a cut-off at overall wave reproducibility 55% or overall response level 4 dB SPL. The prevalence of TEOAEs in left ears was 55.6%. No TEOAEs were found in subjects with a pure-tone average (PTA) above 40 dB HL. In the subgroup with TEOAEs, a significant decrease in overall wave reproducibility with age was found. We compared 45 normal-hearing elderly subjects with TEOAEs with a control group of 20 normal-hearing young adults The elderly had significantly lower mean overall response levels and mean overall wave reproducibility. Average hearing level was significantly higher in the elderly than in controls We conclude that the prevalence of TEOAEs decreases with age, and that the overall response level and overall reproducibility decrease with age. This decrease may not be seen in isolation from the increase in hearing threshold level.

A traditional Thai medicinal extract from Cassia siamea was evaluated with respect to its larvicidal properties by determining the median lethal concentration (LC50) at 24, 48, 72 and 96 h against the fourth instar larvae of Culex quinquefasciatus, which is a carrier of mosquito-borne diseases, by studying the histopathological alterations. The 24, 48, 72 and 96 h LC50 values were 394.29, 350.24, 319.17 and 272.42 ppm, respectively. The histopathological lesions after exposure to 25% of the 24-h LC50 were observed primarily in the midgut of the larva. Lesions with edema, swelling, and deformation or elongation of the epithelial cells were observed. Moreover, cells protruding into the lumen and absent microvilli were also found in some areas. The present study reveals that aqueous C. siamea leaf extracts have natural biopesticide properties. PMID:26868707

Out of seven fungal species belonging to four genera isolated from pond and wallow soils using feathers of Pavo cristatus as bait, four species viz., Aspergillus flavus, Aspergillus niger, Chrysosporium pseudomerdarium and Trichophyton ajelloi were most frequent. Chrysosporium and Trichophyton spp. were more pathogenic on Culex quinquefasciatus larvae than Aspergillus and Penicillium. The bioefficacy tests conducted as per the protocol of World Health Organization and the LC(50) values calculated by the Probit analysis showed that 3(rd)-instar C. quinquefasciatus were more susceptible to the conidia of above fungi. Highest mortality was observed in the larvae of C. quinquefasciatus when exposed to T. ajelloi. The density of fungal conidia was greatest on the ventral brush, palmate hair and anal region of the mosquito larvae after exposing for 72 hours. The potentiality of these fungi for use in the control of C. quinquefasciatus is discussed which can be exploited as a suitable biocontrol agent in the tropics.

This study investigated the toxic effects of essential oils isolated from Siparuna guianensis against Aedes aegypti, Culex quinquefasciatus (eggs, larvae, pupae, and adult) and Aedes albopictus (C6/36) cells. The oviposition-deterring activity, egg viability, and repellence activity in the presence of different essential oils concentrations were determined. The essential oils showed high toxicity to all developmental stages of A. aegypti and C. quinquefasciatus. Furthermore, the oils also showed high repellent activity towards the adult stage of mosquitoes (0.025 to 0.550 μg/cm2 skin conferred 100% repellence up to 120 min) and in contact with cultured insect cells (C6/36) induced death possibly by necrosis. The results presented in this work show the potential of S. guianensis essential oils for the development of an alternative and effective method for the natural control of mosquitoes in homes and urban areas. PMID:25646797

Since ancient times, plant products were used in various aspects. However, their use against pests decreased when chemical products became developed. Recently, concerns increased with respect to public health and environmental security requiring detection of natural products that may be used against insect pests. In this study, 41 plant extracts and 11 oil mixtures were evaluated against the yellow fever mosquito, Aedes aegypti (Linnaeus), the malaria vector, Anopheles stephensi (Liston), and the filariasis and encephalitis vector, Culex quinquefasciatus (Say) (Diptera: Culicidae) using the skin of human volunteers to find out the protection time and repellency. The five most effective oils were those of Litsea (Litsea cubeba), Cajeput (Melaleuca leucadendron), Niaouli (Melaleuca quinquenervia), Violet (Viola odorata), and Catnip (Nepeta cataria), which induced a protection time of 8 h at the maximum and a 100% repellency against all three species. This effect needs, however, a peculiar formulation to fix them on the human skin.

Larvicidal activity of partially purified extracts of leaves of Vitex negundo, Nerium oleander and seeds of Syzygium jambolanum on different instars of Culex quinquefasciatus and Anopheles stephensi was estimated. Petroleum ether (PE): Ethyl acetate (EA) 3:1 fraction of V. negundo, 1:1 fractions of N. oleander and S. jambolanum inflicted considerable larval mortality and interfered with pupal-adult metamorphosis. At very low concentration the active fractions of these plant extracts extended the duration of the various larval instars and of pupation. In general, I and II instar larvae were more susceptible to the active fractions. Species and stage specific differences in the susceptibility of the mosquitoes to the active fractions of the plant extracts were observed.

Pakistan is blessed with plants of Asteraceae family with known medicinal background used for centuries by Hakims (traditional physicians). Keeping in mind the background of their anti-larval potential, a total of 21 endophytic actinomycetes were isolated from four Asteraceae plants and screened against the first and fourth instar stages of Culex quinquefasciatus Say mosquito larvae. Of the 21 isolates, 6 of them gave strong larvicidal activity (80-100% mortality) in the screening results and 4 isolates gave a potent larvicidal activity (100% mortality) at the fourth instar stage. These isolates belonged to different species within the actinomycetes group, namely Streptomyces albovinaceus and Streptomyces badius. This communication reports the larvicidal potential of endophytic actinomycetes residing within the native Asteraceae plants in Pakistan. The study suggests further exploration through large-scale productions leading to the identification of the larvicidal compounds.

For two metropolitan New Jersey counties, monthly average adult mosquito (Culex) catch from New Jersey light trap data sets covering multiple decades is related to a number of meteorological factors. From June through August climatological conditions accounted for between 40% and 50% of the variation in average catch. In general, high monthly precipitation totals both in the month corresponding to the catch and the previous month were associated with increased trap catch. However, individual heavy rainfall events tended to reduce catch. Warm temperatures exerted a positive influence on mosquito abundance in June, but were associated with a low catch in August. Linear meteorological relationships explained only a small percentage of the variations in mosquito catch during May and September. During July, and particularly August, antecedent monthly catch also explained a significant portion of the variance in the contemporaneous catch. Over 60% of the variability in August catch could be attributed to the July population.

Reptiles and amphibians constitute a significant portion of vertebrate biomass in terrestrial ecosystems and may be important arbovirus reservoirs. To investigate mosquito preference for ectothermic hosts, feeding indices were calculated from data collected in Tuskegee National Forest, Alabama, USA. Four mosquito species fed upon ectothermic hosts, with Cx. peccator and Cx. territans feeding primarily upon ectotherms. These two species appeared to target distinct species with little overlap in host choice. Culex peccator was a generalist in its feeding patterns within ectotherms, while Cx. territans appeared to be a more specialized feeder. Six of eleven ectotherm species fed upon by Cx. territans were fed upon more often than predicted based upon abundance. Spring Peepers were highly preferred over other host species by Cx. territans. Blood meals taken from each host species varied temporally, with some hosts being targeted fairly evenly throughout the season and others being fed upon in seasonal peaks. PMID:18981528

Host blood meals in seven mosquito species previously shown to be infected with eastern equine encephalitis virus at a site in the Tuskegee National Forest in southcentral Alabama were investigated. Of 1374 blood meals derived from 88 different host species collected over 6 years from these seven mosquito species, 1099 were derived from Culex erraticus. Analysis of the temporal pattern of Cx. erraticus meals using a Runs test revealed that the patterns of feeding upon avian and mammalian hosts from March to September of each year were not randomly distributed over time. Similarly, meals taken from the three most commonly targeted host species (yellow-crowned night heron, great blue heron, and white-tailed deer) were not randomly distributed. A Tukey's two-way analysis of variance test demonstrated that although the temporal pattern of meals taken from avian hosts were consistent over the years, the patterns of meals taken from the individual host species were not consistent from year to year.

The ability of many insects to learn has been documented. However, a limited number of studies examining associative learning in medically important arthropods has been published. Investigations into the associative learning capabilities of Culex quinquefasciatus Say were conducted by adapting methods commonly used in experiments involving Hymenoptera. Male and female mosquitoes were able to learn a conditioned stimulus that consisted of an odor not normally encountered in nature (synthetic strawberry or vanilla extracts) in association with an unconditioned stimulus consisting of either a sugar (males and females) or blood (females) meal. Such information could lead to a better understanding of the ability of mosquitoes to locate and select host and food resources in nature.

The need to have an improved knowledge on the bioecology of Culex quinquefasciatus, a prerequisite in the development of cost-effective control strategies, has informed the present preliminary investigation to put in better perspective variations that exist in the egg rafts of the species. Freshly laid egg rafts were collected and incubated at ambient temperature in well-labeled plastic trays. The results showed overall inconsistency in all indices monitored for the egg rafts. Generally, survivorship was high for the species. All immature stage and adult parameters measured varied significantly among the egg rafts and between/within sexes of the species. Therefore, this study suggests the presence of inherent variation in the bionomics of egg rafts of C. quinquefasciatus, probably influenced by the environment and hence underscores the need for additional studies to further elucidate the roles of genetics and environment in vectorial competence of the species, in order to develop robust sustainable mosquito vector control protocols. PMID:27013900

Enviromental conditions, including such important climatic variables as temperature and precipitation, change with altitude; thus, elevation plays a significant role in determining population and community structure in a variety of organisms. Using single nucleotide polymorphisms (SNPs) and geometric morphometrics, nine populations of Culex theileri Theobald occurring in different ecological subregions at altitudes between 808-2,130 m in northeastern Turkey were compared. The wing size and shape data indicate that there are significant phenotypic differences among them, while Cx theileri populations are not genetically differentiated in the northeast part of Turkey. The size and shape variation analysis of wings showed that there is a positive correlation between wing (body) size/shape and altitude.

Background The Americas have suffered a dramatic epidemic of Zika since May in 2015, when Zika virus (ZIKV) was first detected in Brazil. Mosquitoes belonging to subgenus Stegomyia of Aedes, particularly Aedes aegypti, are considered the primary vectors of ZIKV. However, the rapid spread of the virus across the continent raised several concerns about the transmission dynamics, especially about potential mosquito vectors. The purpose of this work was to assess the vector competence of the house mosquito Culex quinquefasciatus from an epidemic Zika area, Rio de Janeiro, Brazil, for local circulating ZIKV isolates. Methodology/Principal Findings Culex quinquefasciatus and Ae. aegypti (positive control of ZIKV infection) from Rio de Janeiro were orally exposed to two ZIKV strains isolated from human cases from Rio de Janeiro (Rio-U1 and Rio-S1). Fully engorged mosquitoes were held in incubators at 26 ± 1°C, 12 h:12 h light:dark cycle and 70 ± 10% humidity. For each combination mosquito population—ZIKV strain, 30 specimens were examined for infection, dissemination and transmission rates, at 7, 14 and 21 days after virus exposure by analyzing body (thorax plus abdomen), head and saliva respectively. Infection rates were minimal to completely absent in all Cx. quinquefasciatus-virus combinations and were significantly high for Ae. aegypti. Moreover, dissemination and transmission were not detected in any Cx. quinquefasciatus mosquitoes whatever the incubation period and the ZIKV isolate. In contrast, Ae. aegypti ensured high viral dissemination and moderate to very high transmission. Conclusions/Significance The southern house mosquito Cx. quinquefasciatus from Rio de Janeiro was not competent to transmit local strains of ZIKV. Thus, there is no experimental evidence that Cx. quinquefasciatus likely plays a role in the ZIKV transmission. Consequently, at least in Rio, mosquito control to reduce ZIKV transmission should remain focused on Ae. aegypti. PMID

In Culex quinquefasciatus, CquiOR91 is the ortholog of 2 larvae-specific odorant receptors (ORs) from Anopheles gambiae (Agam\\Or40, previously shown to respond to several odorant ligands including the broad-spectrum repellent N,N-diethyl-3-methylbenzamide, DEET) and Aedes aegypti (Aaeg\\Or40). When we cloned full-length CquiOR91 from a Culex quinquefasciatus larval head RNA sample, we found 2 alleles of this OR, differing at 9 residues. Functional analysis using the Xenopus oocyte expression system and 2-electrode voltage clamp electrophysiology revealed one allele (CquiOR91.1) to be nonfunctional, whereas the other allele (CquiOR91.2) was functional. Receptors formed by CquiOR91.2 and Cqui\\Orco responded to (-)-fenchone, (+)-fenchone, and DEET, similar to what has been reported for Agam\\Or40. We also identified 5 novel odorant ligands for the CquiOR91.2 + Cqui\\Orco receptor: 2-isobutylthiazole, veratrole, eucalyptol, d-camphor, and safranal, with safranal being the most potent. To explore possible reasons for the lack of function for CquiOR91.1, we generated a series of mutant CquiOR91.2 subunits, in which the residue at each of the 9 polymorphic residue positions was changed from what occurs in CquiOR91.2 to what occurs in CquiOR91.1. Eight of the 9 mutant versions of CquiOR91.2 formed functional receptors, responding to (-)-fenchone. Only the CquiOR91.2 Y183C mutant was nonfunctional. The reverse mutation (C183Y) conferred function on CquiOR91.1 , which became responsive to (-)-fenchone and safranal. These results indicate that the "defect" in CquiOR91.1 that prevents function is the cysteine at position 183.

Abstract Ornithophilic Culex species are considered the primary amplification vectors of West Nile virus (WNV) in bird hosts as well as vectors responsible for epidemic transmission. Culex coronator was first collected from Okaloosa, Santa Rosa, Walton, and Washington Counties in Florida in 2005 and has since spread throughout the state. The vector competence of Cx. coronator for WNV, known to be infected in nature, has not been assessed. Without this knowledge, we are unable to assess this species' potential as an enzootic and epidemic vector of WNV in Florida. In the current study, we investigate the reproductive biology and susceptibility to WNV infection, dissemination, and transmission by Cx. coronator. We show that Cx. coronator is capable of delaying oviposition for several weeks after blood feeding and that the number of eggs laid is greater for avian than mammalian hosts. Cx. coronator were highly susceptible to infection (∼80–100%) and dissemination (∼65–85% by 18 days since exposure) with lower rates of transmission (0–17% at 25°C and 28–67% at 28°C), suggesting that it is a competent vector of WNV under some conditions. The proportion of mosquitoes with disseminated infections related to the time since exposure and was higher at 28°C than at 25°C. The rapid and statewide distribution of Cx. coronator throughout Florida poses as a potential public health risk. This baseline knowledge is essential information for mosquito control and public health agencies to assess current and future disease risk to Southeastern United States. PMID:25072992

Distortion product otoacoustic emissions (DPOAEs) are traditionally evoked by two-tone stimuli. In this study, emission data from Mongolian gerbils are reported that were obtained with stimuli consisting of six to 10 tones. The stimuli were constructed by replacing one of the tones of a tone pair by a narrowband multitone complex. This produced rich spectra of the ear canal sound pressure in which many of the third-order DPOAEs originated from the interaction of triplets of stimulus components. A careful choice of the stimulus frequencies ensured that none of these DPOAE components coincided. Three groups of DPOAEs are reported, two of which are closely related to DPOAEs evoked by tone pairs. The third group has no two-tone equivalent and only arises when using a multitone stimulus. We analyzed the relation between multitone-evoked DPOAEs and DPOAEs evoked by tone pairs, and explored the new degrees of freedom offered by the multitone paradigm. PMID:20838846

Background To characterize the findings of brainstem auditory evoked potential in HIV-positive individuals exposed and not exposed to antiretroviral treatment. Material/Methods This research was a cross-sectional, observational, and descriptive study. Forty-five HIV-positive individuals (18 not exposed and 27 exposed to the antiretroviral treatment – research groups I and II, respectively – and 30 control group individuals) were assessed through brainstem auditory evoked potential. Results There were no significant between-group differences regarding wave latencies. A higher percentage of altered brainstem auditory evoked potential was observed in the HIV-positive groups when compared to the control group. The most common alteration was in the low brainstem. Conclusions HIV-positive individuals have a higher percentage of altered brainstem auditory evoked potential that suggests central auditory pathway impairment when compared to HIV-negative individuals. There was no significant difference between individuals exposed and not exposed to antiretroviral treatment. PMID:26485202

This study measured the postural sway and brain potentials evoked by a visual depth stimulus. Thirteen subjects maintained standing posture on a force platform, and were administered two types of depth stimuli, strong and weak. The latency and amplitude of evoked potentials as well as changes in center of foot pressure (CFP) and the electromyogram (EMG) were examined. CFP displacement was found to change according to stimulus intensity. In the occipital lobe, evoked potentials exhibited a triphasic peak, with the first positive peak at approximately 120 ms (P120), the first negative peak at approximately 160 ms (N200), and the second positive peak at approximately 260 ms (P250). Brain evoked potentials correlated with CFP displacement as well as the latency of onset of EMG response. Onset of EMG response was probably related to the P120 component, whereas CFP displacement was related to the P250 component.

Music is capable of evoking exceptionally strong emotions and of reliably affecting the mood of individuals. Functional neuroimaging and lesion studies show that music-evoked emotions can modulate activity in virtually all limbic and paralimbic brain structures. These structures are crucially involved in the initiation, generation, detection, maintenance, regulation and termination of emotions that have survival value for the individual and the species. Therefore, at least some music-evoked emotions involve the very core of evolutionarily adaptive neuroaffective mechanisms. Because dysfunctions in these structures are related to emotional disorders, a better understanding of music-evoked emotions and their neural correlates can lead to a more systematic and effective use of music in therapy.

Several approaches, based on different assumptions and with various degree of theoretical sophistication and implementation complexity, have been developed for improving the measurement of evoked potentials (EP) performed by conventional averaging (CA). In many of these methods, one of the major challenges is the exploitation of a priori knowledge. In this paper, we present a new method where the 2nd-order statistical information on the background EEG and on the unknown EP, necessary for the optimal filtering of each sweep in a Bayesian estimation framework, is, respectively, estimated from pre-stimulus data and obtained through a multiple integration of a white noise process model. The latter model is flexible (i.e. it can be employed for a large class of EP) and simple enough to be easily identifiable from the post-stimulus data thanks to a smoothing criterion. The mean EP is determined as the weighted average of the filtered sweeps, where each weight is inversely proportional to the expected value of the norm of the correspondent filter error, a quantity determinable thanks to the employment of the Bayesian approach. The performance of the new approach is shown on both simulated and real auditory EP. A signal-to-noise ratio enhancement is obtained that can allow the (possibly automatic) identification of peak latencies and amplitudes with less sweeps than those required by CA. For cochlear EP, the method also allows the audiology investigator to gather new and clinically important information. The possibility of handling single-sweep analysis with further development of the method is also addressed.

MULTIPLE COLOR STIMULUS INDUCED STEADY STATE VISUAL EVOKED POTENTIALS M. Cheng, X. Gao, S. Gao, D. Xu Institute of Biomedical Engineering...characteristics of high SNR and effectiveness in short-term identification of evoked responses. In most of the SSVEP experiments, single high...frequency stimuli are used. To characterize the complex rhythms in SSVEP, a new multiple color stimulus pattern is proposed in this paper. FFT and

The central nucleus of the inferior colliculus (ICc) receives a large number of convergent inputs that are both excitatory and inhibitory. Although excitatory inputs typically are evoked by stimulation of the contralateral ear, inhibitory inputs can be recruited by either ear. Here we evaluate ipsilaterally evoked inhibition in single ICc cells in awake Mexican free-tailed bats. The principal question we addressed concerns the degree to which ipsilateral inhibition at the ICc suppresses contralaterally evoked discharges and thus creates the excitatory-inhibitory (EI) properties of ICc neurons. To study ipsilaterally evoked inhibition, we iontophoretically applied excitatory neurotransmitters and visualized the ipsilateral inhibition as a gap in the carpet of background activity evoked by the transmitters. Ipsilateral inhibition was seen in 86% of ICc cells. The inhibition in most cells had both glycinergic and GABAergic components that could be blocked by the iontophoretic application of bicuculline and strychnine. In 80% of the cells that were inhibited, the ipsilateral inhibition and contralateral excitation were temporally coincident. In many of these cells, the ipsilateral inhibition suppressed contralateral discharges and thus generated the cell's EI property in the ICc. In other cells, the ipsilateral inhibition was coincident with the initial portion of the excitation, but the inhibition was only 2-4 ms in duration and suppressed only the first few contralaterally evoked discharges. The suppression was so slight that it often could not be detected as a decrease in the spike count generated by increasing ipsilateral intensities. Twenty percent of the cells that expressed inhibition, however, had inhibitory latencies that were longer than the excitatory latencies. In these neurons, the inhibition arrived too late to suppress most or any of the discharges. Finally, in the majority of cells, the ipsilateral inhibition persisted for tens of milliseconds beyond

The phenomenon in which a certain smell evokes a specific memory is known as the Proust phenomenon. Odor-evoked autobiographic memories are more emotional than those elicited by other sensory stimuli. The results of our previous study indicated that odor-evoked autobiographic memory accompanied by positive emotions has remarkable effects on various psychological and physiological activities, including the secretion of cytokines, which are immune-signaling molecules that modulate systemic inflammation. In this study, we aimed to clarify the neural substrates associated with the interaction between odor-evoked autobiographic memory and peripheral circulating cytokines. We recruited healthy male and female volunteers and investigated the association between brain responses and the concentration of several cytokines in the plasma by using positron emission tomography (PET) recordings when an autographic memory was evoked in participants by asking them to smell an odor that was nostalgic to them. Participants experienced positive emotions and autobiographic memories when nostalgic odors were presented to them. The levels of peripheral proinflammatory cytokines, such as the tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), were significantly reduced after experiencing odor-evoked autobiographic memory. Subtraction analysis of PET images indicated that the medial orbitofrontal cortex (mOFC) and precuneus/posterior cingulate cortex (PCC) were significantly activated during experiences of odor-evoked autobiographic memory. Furthermore, a correlation analysis indicated that activities of the mOFC and precuneus/PCC were negatively correlated with IFN-γ concentration. These results indicate that the neural networks including the precuneus/PCC and mOFC might regulate the secretion of peripheral proinflammatory cytokines during the experience of odor-evoked autobiographic memories accompanied with positive emotions. PMID:23977312

Even though disgust and fear are both negative emotions, they are characterized by different physiology and action tendencies. The aim of this study was to examine whether fear- and disgust-evoking images would produce different attention bias effects, specifically those related to attention (dis)engagement. Participants were asked to identify a target which was briefly presented around a central image cue, which could either be disgusting, frightening, or neutral. The interval between cue onset and target presentation varied within blocks (200, 500, 800, 1100 ms), allowing us to investigate the time course of attention engagement. Accuracy was lower and reaction times were longer when targets quickly (200 ms) followed disgust-evoking images than when they followed neutral- or fear-evoking images. For the other, longer interval conditions no significant image effects were found. These results suggest that emotion-specific attention effects can be found at very early visual processing stages and that only disgust-evoking images, and not fear-evoking ones, keep hold of our attention for longer. We speculate that this increase in early attention allocation is related to the need to perform a more comprehensive risk-assessment of the disgust-evoking images. The outcomes underline not only the importance of examining the time course of emotion induced attention effects but also the need to look beyond the dimensions of valence and arousal.

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. However, the use of synthetic insecticides to control Culicidae may lead to resistance, high operational costs and adverse non-target effects. Nowadays, plant-borne mosquitocides may serve as suitable alternative in the fight against mosquito vectors. In this study, the mosquito larvicidal activity of Origanum vulgare (Lamiaceae) leaf essential oil (EO) and its major chemical constituents was evaluated against the malaria vectors Anopheles stephensi and An. subpictus, the filariasis vector Culex quinquefasciatus and the Japanese encephalitis vector Cx. tritaeniorhynchus. The chemical composition of the EO was analyzed by gas chromatography-mass spectroscopy. GC-MS revealed that the essential oil of O. vulgare contained 17 compounds. The major chemical components were carvacrol (38.30%) and terpinen-4-ol (28.70%). EO had a significant toxic effect against early third-stage larvae of An. stephensi, An. subpictus, Cx. quinquefasciatus and Cx. tritaeniorhynchus, with LC50 values of 67.00, 74.14, 80.35 and 84.93 μg/ml. The two major constituents extracted from the O. vulgare EO were tested individually for acute toxicity against larvae of the four mosquito vectors. Carvacrol and terpinen-4-ol appeared to be most effective against An. stephensi (LC50=21.15 and 43.27 μg/ml, respectively) followed by An. subpictus (LC50=24.06 and 47.73 μg/ml), Cx. quinquefasciatus (LC50=26.08 and 52.19 μg/ml) and Cx. tritaeniorhynchus (LC50=27.95 and 54.87 μg/ml). Overall, this research adds knowledge to develop newer and safer natural larvicides against malaria, filariasis and Japanese encephalitis mosquito vectors.

Insect-specific viruses belonging to significant arboviral families have recently been discovered. These viruses appear to be maintained within the insect population without the requirement for replication in a vertebrate host. Mosquitoes collected from Badu Island in the Torres Strait in 2003 were analysed for insect-specific viruses. A novel bunyavirus was isolated in high prevalence from Culex spp. The new virus, provisionally called Badu virus (BADUV), replicated in mosquito cells of both Culex and Aedes origin, but failed to replicate in vertebrate cells. Genomic sequencing revealed that the virus was distinct from sequenced bunyavirus isolates reported to date, but phylogenetically clustered most closely with recently discovered mosquito-borne, insect-specific bunyaviruses in the newly proposed Goukovirus genus. The detection of a functional furin cleavage motif upstream of the two glycoproteins in the M segment-encoded polyprotein suggests that BADUV may employ a unique strategy to process the virion glycoproteins.

A laboratory investigation was undertaken to study the cyclic usage of field recommended doses of Bacillus thuringiensis israelensis (Bti), Bacillus sphaericus (Bsp) and combination of Bti and Bsp (half the recommended dose of each) with deltamethrin 2.8 EC to attain better control of mosquito larvae. To understand the susceptibility status of the Culex quinquefasciatus larvae in the field, early fourth instar field collected larvae were exposed to the above treatments of biopesticides and were monitored for their persistency till 50% mortality was obtained. The larvae those survived in each treatment were transferred to the concentration of deltamethrin that gave 50% mortality. Observations on larval mortality after the treatment with deltamethrin were recorded after 24 h. The results revealed that Bti excels Bsp, as it recorded 54% mortality only on 17th day after application. The other salient finding of this study is LC50 of deltamethrin is sufficient to follow the biopesticides application for an effective control of Culex larvae.

Culex (Acallyntrum) miyagii is described as a new species from Seram Island, Indonesia. The adult male, female, pupa, and larva are described in detail, and illustrations of the male genitalia, pupa, and larva are provided. Cx. miyagii is a forest species breeding in inflorescences of Costus sp. (Costaceae). Keys are provided for identifying the females and 4th-instar larvae of the species of Acallyntrum.

Bioinsecticide Bacillus thuringiensis var. kurstaki (Btk) was used for controlling the mosquito species (Anopheles stephensi and Culex quinquefasciatus) which gave a significant (p<0.05) mortality in both species. The higher concentration of Btk was highly effective compared to the control ones. The controlling effect was dose and time dependent. Among the studied mosquitoes the C. quinquefasciatus (LC5 0.154%) was more susceptible than A. stephensi (LC50 0.372%) towards the bioinsecticide Btk.

Adult Culex quinquefasciatus, maintained from emergence on sugar, were fed blood and then fed either sugar (control) or water (starving) for 7 days. Analysis of ovaries and egg rafts for protein, lipids and glycogen showed that only glycogen levels were diminished by starvation. Eggs from both control and starving females, however, were equally viable. Nonbloodfed starving females lived longer than bloodfed starving females. These results suggest that the blood meal maximizes fertility, not longevity.

This study explores listeners' experience of music-evoked sadness. Sadness is typically assumed to be undesirable and is therefore usually avoided in everyday life. Yet the question remains: Why do people seek and appreciate sadness in music? We present findings from an online survey with both Western and Eastern participants (N = 772). The survey investigates the rewarding aspects of music-evoked sadness, as well as the relative contribution of listener characteristics and situational factors to the appreciation of sad music. The survey also examines the different principles through which sadness is evoked by music, and their interaction with personality traits. Results show 4 different rewards of music-evoked sadness: reward of imagination, emotion regulation, empathy, and no "real-life" implications. Moreover, appreciation of sad music follows a mood-congruent fashion and is greater among individuals with high empathy and low emotional stability. Surprisingly, nostalgia rather than sadness is the most frequent emotion evoked by sad music. Correspondingly, memory was rated as the most important principle through which sadness is evoked. Finally, the trait empathy contributes to the evocation of sadness via contagion, appraisal, and by engaging social functions. The present findings indicate that emotional responses to sad music are multifaceted, are modulated by empathy, and are linked with a multidimensional experience of pleasure. These results were corroborated by a follow-up survey on happy music, which indicated differences between the emotional experiences resulting from listening to sad versus happy music. This is the first comprehensive survey of music-evoked sadness, revealing that listening to sad music can lead to beneficial emotional effects such as regulation of negative emotion and mood as well as consolation. Such beneficial emotional effects constitute the prime motivations for engaging with sad music in everyday life.

This study explores listeners’ experience of music-evoked sadness. Sadness is typically assumed to be undesirable and is therefore usually avoided in everyday life. Yet the question remains: Why do people seek and appreciate sadness in music? We present findings from an online survey with both Western and Eastern participants (N = 772). The survey investigates the rewarding aspects of music-evoked sadness, as well as the relative contribution of listener characteristics and situational factors to the appreciation of sad music. The survey also examines the different principles through which sadness is evoked by music, and their interaction with personality traits. Results show 4 different rewards of music-evoked sadness: reward of imagination, emotion regulation, empathy, and no “real-life” implications. Moreover, appreciation of sad music follows a mood-congruent fashion and is greater among individuals with high empathy and low emotional stability. Surprisingly, nostalgia rather than sadness is the most frequent emotion evoked by sad music. Correspondingly, memory was rated as the most important principle through which sadness is evoked. Finally, the trait empathy contributes to the evocation of sadness via contagion, appraisal, and by engaging social functions. The present findings indicate that emotional responses to sad music are multifaceted, are modulated by empathy, and are linked with a multidimensional experience of pleasure. These results were corroborated by a follow-up survey on happy music, which indicated differences between the emotional experiences resulting from listening to sad versus happy music. This is the first comprehensive survey of music-evoked sadness, revealing that listening to sad music can lead to beneficial emotional effects such as regulation of negative emotion and mood as well as consolation. Such beneficial emotional effects constitute the prime motivations for engaging with sad music in everyday life. PMID:25330315

The aim of this study was to examine the collision responsible for the volitional V-wave evoked by supramaximal electrical stimulation of the motor nerve during voluntary contraction. V-wave was conditioned by transcranial magnetic stimulation (TMS) over the motor cortex at several inter-stimuli intervals (ISI) during weak voluntary plantar flexions (n = 10) and at rest for flexor carpi radialis muscle (FCR; n = 6). Conditioning stimulations were induced by TMS with intensity eliciting maximal motor-evoked potential (MEPmax). ISIs used were ranging from -20 to +20 msec depending on muscles tested. The results showed that, for triceps surae muscles, conditioning TMS increased the V-wave amplitude (~ +250%) and the associated mechanical response (~ +30%) during weak voluntary plantar flexion (10% of the maximal voluntary contraction -MVC) for ISIs ranging from +6 to +18 msec. Similar effect was observed at rest for the FCR with ISI ranging from +6 to +12 msec. When the level of force was increased from 10 to 50% MVC or the conditioning TMS intensity was reduced to elicit responses of 50% of MEPmax, a significant decrease in the conditioned V-wave amplitude was observed for the triceps surae muscles, linearly correlated to the changes in MEP amplitude. The slope of this correlation, as well as the electro-mechanical efficiency, was closed to the identity line, indicating that V-wave impact at muscle level seems to be similar to the impact of cortical stimulation. All these results suggest that change in V-wave amplitude is a great index to reflect changes in cortical neural drive addressed to spinal motoneurons.

Measurements of brainstem auditory evoked potentials (BAEP) and middle latency auditory evoked potentials (MLAEP) are readily available neurophysiologic assessments. The generators for BAEP are believed to involve the structures of cochlear nerve, cochlear nucleus, superior olive complex, dorsal and rostral pons, and lateral lemniscus. The generators for MLAEP are assumed to be located in the subcortical area and auditory cortex. BAEP are commonly used in evaluating children with autistic and hearing disorders. However, measurement of MLAEP is rarely performed in young children. To explore the feasibility of this procedure in young children, we retrospectively reviewed our neurophysiology databank and charts for a 3-year period to identify subjects who had both BAEP and MLAEP performed. Subjects with known or identifiable central nervous system abnormalities from the history, neurologic examination and neuroimaging studies were excluded. This cohort of 93 children up to 3 years of age was divided into 10 groups based on the age at testing (upper limits of: 1 week; 1, 2, 4, 6, 8, 10 and 12 months; 2 years; and 3 years of age). Evolution of peak latency, interpeak latency and amplitude of waveforms in BAEP and MLAEP were demonstrated. We concluded that measurement of BAEP and MLAEP is feasible in children, as early as the first few months of life. The combination of both MLAEP and BAEP may increase the diagnostic sensitivity of neurophysiologic assessment of the integrity or functional status of both the peripheral (acoustic nerve) and the central (brainstem, subcortical and cortical) auditory conduction systems in young children with developmental speech and language disorders.

Field studies were conducted in a rice, Oryza sativa L., agroecosystem in Mwea Kenya to compare the efficiency of CO2-baited Centers for Disease Control (CDC) light traps against nonbaited CDC light traps and gravid traps against oviposition traps in outdoor collection of Culex quinquefasciatus Say (Diptera: Culicidae) and other mosquitoes. Collectively, 21 mosquito species from the genera Culex, Anopheles, Mansonia, Ficalbia, and Aedes were captured during the 10-wk study period. Cx. quinquefasciatus was the predominant species in all trap types with proportions ranging from 57% in the nonbaited CDC light traps to 95% in the gravid traps. Significantly higher numbers of Cx. quinquefasciatus and Culex annulioris Theobald were collected in the CO2-baited CDC light traps than in the nonbaited CDC light traps, but the numbers of other mosquito species, including malaria vectors Anopheles arabiensis Patton and Anopheles funestus Giles did not differ significantly between the two trap types. More Cx. quinquefasciatus females were collected in grass infusion-baited gravid traps than egg rafts of this species in oviposition traps containing the same infusion. Although most mosquitoes captured in CO,-baited and nonbaited CDC light traps were unfed, most of those collected in gravid traps were gravid. From these findings, it is concluded that at least in the rice-growing area of Mwea Kenya, CO2-baited CDC light traps in conjunction with gravid traps can be used in monitoring of Cx. quinquefasciatus both for control and disease surveillance.

An initiative within the US military is targeting the replacement of traditional munitions constituents with insensitive munitions to reduce risk of accidental detonation. The purpose of the present study was to comparatively assess toxicity of the traditional munitions constituents 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) with the new insensitive munitions constituents 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO). The following exposure durations were performed with Rana pipiens (leopard frog) tadpoles: TNT and DNAN, 96 h and 28 d; RDX, 10 d and 28 d; NTO, 28 d. The 96-h 50% lethal concentration (LC50) values and 95% confidence intervals for TNT and DNAN were 4.4 mg/L (4.2 mg/L, 4. 7 mg/L) and 24.3 mg/L (21.3 mg/L, 27.6 mg/L), respectively. No significant impacts on survival were observed in the 10-d exposure to RDX up to 25.3 mg/L. Effects on tadpole swimming distance were observed with a lowest-observed-effect concentration (LOEC) of 5.9 mg/L RDX. In the 28-d exposures, the LOECs for survival for TNT, DNAN, and NTO were 0.003 mg/L, 2.4 mg/L, and 5.0 mg/L, respectively. No significant mortality was observed in the RDX chronic 28-d exposure up to the highest treatment level tested of 28.0 mg/L. Neither tadpole developmental stage nor growth was significantly affected in any of the 28-d exposures. Rana pipiens were very sensitive to chronic TNT exposure, with an LOEC 3 orders of magnitude lower than those for insensitive munitions constituents DNAN and NTO.

Visual information is fundamental to how we appreciate our environment and interact with others. The visual evoked potential (VEP) is among those evoked potentials that are the bioelectric signals generated in the striate and extrastriate cortex when the retina is stimulated with light which can be recorded from the scalp electrodes. In the current paper, we provide an overview of the various modalities, techniques, and methodologies which have been employed for visual evoked potentials over the years. In the first part of the paper, we cast a cursory glance on the historical aspect of evoked potentials. Then the growing clinical significance and advantages of VEPs in clinical disorders have been briefly described, followed by the discussion on the earlier and currently available methods for VEPs based on the studies in the past and recent times. Next, we mention the standards and protocols laid down by the authorized agencies. We then summarize the recently developed techniques for VEP. In the concluding section, we lay down prospective research directives related to fundamental and applied aspects of VEPs as well as offering perspectives for further research to stimulate inquiry into the role of visual evoked potentials in visual processing impairment related disorders. PMID:27034907

Acoustic sensitivity of the vestibular apparatus is well-established, but the contribution of vestibular receptors to the late auditory evoked potentials of cortical origin is unknown. Evoked potentials from 500 Hz tone pips were recorded using 70 channel EEG at several intensities below and above the vestibular acoustic threshold, as determined by vestibular evoked myogenic potentials (VEMPs). In healthy subjects both auditory mid- and long-latency auditory evoked potentials (AEPs), consisting of Na, Pa, N1 and P2 waves, were observed in the sub-threshold conditions. However, in passing through the vestibular threshold, systematic changes were observed in the morphology of the potentials and in the intensity dependence of their amplitude and latency. These changes were absent in a patient without functioning vestibular receptors. In particular, for the healthy subjects there was a fronto-central negativity, which appeared at about 42 ms, referred to as an N42, prior to the AEP N1. Source analysis of both the N42 and N1 indicated involvement of cingulate cortex, as well as bilateral superior temporal cortex. Our findings are best explained by vestibular receptors contributing to what were hitherto considered as purely auditory evoked potentials and in addition tentatively identify a new component that appears to be primarily of vestibular origin.

We present a method for the analysis of electroencephalograms (EEG). In particular, small signals due to stimulation, so-called evoked potentials (EPs), have to be detected in the background EEG. This is achieved by using a denoising implementation based on the wavelet decomposition. One recording of visual evoked potentials, and recordings of auditory evoked potentials from four subjects corresponding to different age groups are analyzed. We find higher variability in older individuals. Moreover, since the EPs are identified at the single stimulus level (without need of ensemble averaging), this will allow the calculation of better resolved averages. Since the method is parameter free (i.e. it does not need to be adapted to the particular characteristics of each recording), implementations in clinical settings are imaginable.

Confirmation is provided for the belief that evoked potentials may reflect differences in hemispheric functioning that are marginal at best. Subjects were right-handed and audiologically normal men and women, and responses were recorded using standard EEG techniques. Subjects were instructed to listen for the targets while laying in a darkened sound booth. Different stimuli, speech and tone signals, were used. Speech sounds were shown to evoke a response pattern that resembles that to tone or clicks. Analysis of variances on peak amplitude and latency measures showed no significant differences between hemispheres, however, a Wilcoxon test showed significant differences in hemispheres for certain target tasks.

The authors studied auditory brainstem evoked potentials (BAEP) in 27 organ donors aged 40 to 68 years treated in neurosurgery units in Szczecin and Grenoble. Abnormal results were found in all cases. In 63% of cases no evoked action potentials were obtained, in 34% only the 1st wave was obtained, and in two cases evolution was observed with activity extinction. The authors believe that in the process of shaping of BAEP morphotic extinction begins from the later waves to earlier ones in agreement with the rostrocaudal direction of extinction of the functions or brain midline structures, and in a single study various findings may be obtained.

Motor unit action potentials (MUAPs) evoked by repetitive, low-intensity transcranial magnetic stimulation can be modeled as a Poisson process. A mathematical consequence of such a model is that the ratio of the variance to the mean of the amplitudes of motor evoked potentials (MEPs) should provide an estimate of the mean size of the individual MUAPs that summate to generate each MEP. We found that this is, in fact, the case. Our finding thus supports the use of the Poisson distribution to model MEP generation and indicates that this model enables characterization of the motor unit population that contributes to near-threshold MEPs.

A study is being conducted to explore use of the steady state visual-evoke electrocortical response as an indicator of cognitive task loading. Application of linear descriptive modeling to steady state Visual Evoked Response (VER) data is summarized. Two aspects of linear modeling are reviewed: (1) unwrapping the phase-shift portion of the frequency response, and (2) parsimonious characterization of task-loading effects in terms of changes in model parameters. Model-based phase unwrapping appears to be most reliable in applications, such as manual control, where theoretical models are available. Linear descriptive modeling of the VER has not yet been shown to provide consistent and readily interpretable results.

Auditory potentials were recorded from bipolar electrodes chronically implanted in the cochlear nuclei of four cats. In a training box modified to reduce echoes these animals were exposed to clicks and tone pulses presented from an overhead speaker. Slight changes in the position of the animal in the resulting sound field produced marked changes in the potentials evoked from the cochlear nucleus. These phenomena were observed in the unanesthetized, unrestrained subjects as well as in those under Nembutal anesthesia. It is suggested that these acoustic effects complicate the analysis and interpretation of potentials evoked from the cochlear nucleus under conditions of habituation, shifts in attention, and learning.

The resistance status towards permethrin among the laboratory strain, the permethrin-selected strain and four field strains of Culex quinquefasciatus collected in Kuala Lumpur, Malaysia was determined using three standard laboratory methods: WHO larval bioassay, WHO adult bioassay and biochemical microplate assay. Cx. quinquefasciatus permethrin-selected strain larvae were the least susceptible to permethrin with a resistance ratio of 47.28-folds, whereas all field strain larvae of the same species were tolerant to permethrin with resistance ratios of more than 3-folds. In contrast, in adult stage, the permethrin exposed permethrin-selected strain (resistance ratio = 1.27) was found to be more susceptible to permethrin than all permethrin-exposed field strains (resistance ratios = 2.23–2.48). Complete mortalities for all strains of Cx. quinquefasciatus adults proved the effectiveness of the synergist; piperonyl butoxide (PBO). For the biochemical microplate assay, the reduction of the mean optical density of elevated oxidase activity of three field strains upon exposure to PBO confirmed the association between oxidase activity and permethrin tolerance. On the other hand, irregular patterns of the mean optical density of elevated oxidase activity in the laboratory strain, permethrin-selected strain and Jalan Fletcher strain illustrated the gene variation within these mosquito colonies as well as the involvement of other enzyme activities in the permethrin resistance occurred. PMID:23961241

Introduction Genetic modification, or transgenesis, is a powerful technique to investigate the molecular interactions between vector-borne pathogens and their arthropod hosts, as well as a potential novel approach for vector-borne disease control. Transgenesis requires the use of specific regulatory regions, or promoters, to drive expression of genes of interest in desired target tissues. In mosquitoes, the vast majority of described promoters are from Anopheles and Aedes mosquitoes. Results Culex tarsalis is one of the most important vectors of arboviruses (including West Nile virus) in North America, yet it has not been the subject of molecular genetic study. In order to facilitate molecular genetic work in this important vector species, we isolated four fat body-specific promoter sequences located upstream of the Cx. tarsalis vitellogenin genes (Vg1a, Vg1b, Vg2a and Vg2b). Sequences were analyzed in silico to identify requisite cis-acting elements. The ability for promoter sequences to drive expression of green fluorescent protein (GFP) in vivo was investigated using transgenic Drosophila melanogaster. All four promoters were able to drive GFP expression but there was dramatic variation between promoters and between individual Drosophila lines, indicating significant position effects. The highest expression was observed in line Vg2bL3, which was >300-fold higher than the lowest line Vg1aL2. Conclusions These new promoters will be useful for driving expression of genes of interest in transgenic Cx. tarsalis and perhaps other insects. PMID:24586476

Filariasis is a mosquito-borne disease that causes lymphedema and the main vector is Culex quinquefasciatus. A simple measure was taken to eradicate the vector using nanoemulsion. Eucalyptus oil nanoemulsion was formulated in various ratios comprising of eucalyptus oil, tween 80 and water by ultrasonication. The stability of nanoemulsion was observed over a period of time and 1:2 ratios of eucalyptus oil (6%) and surfactant (12%) was found to be stable. The formulated eucalyptus oil nanoemulsion was characterized by transmission electron microscopy and dynamic light scattering. The nanoemulsion droplets were found to have a Z-average diameter of 9.4 nm and were spherical in shape. The larvicidal activity of eucalyptus oil nanoemulsion and bulk emulsion was tested and compared. Our nanoemulsion showed higher activity when compared to bulk emulsion. The histopathology of larvae-treated and untreated nanoemulsion was analyzed. Furthermore, biochemical assays were carried out to examine the effect of nanoemulsion on biochemical characteristics of larvae. The treated larval homogenate showed decrease in total protein content and a significant reduction in the levels of acetylcholinesterase. The levels of acid and alkaline phosphatase also showed reduction as compared to control larval homogenate.

We describe the isolation and characterization of a novel flavivirus, isolated from a pool of Culex (Melanoconion) ocossa Dyar and Knab mosquitoes collected in 2009 in an urban area of the Amazon basin city of Iquitos, Peru. Flavivirus infection was detected by indirect immunofluorescent assay of inoculated C6/36 cells using polyclonal flavivirus antibodies (St. Louis encephalitis virus, yellow fever virus and dengue virus type 1) and confirmed by RT-PCR. Based on partial sequencing of the E and NS5 gene regions, the virus isolate was most closely related to the mosquito-borne flaviviruses but divergent from known species, with less than 45 and 71 % pairwise amino acid identity in the E and NS5 gene products, respectively. Phylogenetic analysis of E and NS5 amino acid sequences demonstrated that this flavivirus grouped with mosquito-borne flaviviruses, forming a clade with Nounané virus (NOUV). Like NOUV, no replication was detected in a variety of mammalian cells (Vero-76, Vero-E6, BHK, LLCMK, MDCK, A549 and RD) or in intracerebrally inoculated newborn mice. We tentatively designate this genetically distinct flavivirus as representing a novel species, Nanay virus, after the river near where it was first detected.

We present a population model to understand the effects of temperature and rainfall on the population dynamics of the southern house mosquito, Culex quinquefasciatus Say, along an elevational gradient in Hawaii. We use a novel approach to model the effects of temperature on population growth by dynamically incorporating developmental rate into the transition matrix, by using physiological ages of immatures instead of chronological age or stages. We also model the effects of rainfall on survival of immatures as the cumulative number of days below a certain rain threshold. Finally, we incorporate density dependence into the model as competition between immatures within breeding sites. Our model predicts the upper altitudinal distributions of Cx. quinquefasciatus on the Big Island of Hawaii for self-sustaining mosquito and migrating summer sink populations at 1,475 and 1,715 m above sea level, respectively. Our model predicts that mosquitoes at lower elevations can grow under a broader range of rainfall parameters than middle and high elevation populations. Density dependence in conjunction with the seasonal forcing imposed by temperature and rain creates cycles in the dynamics of the population that peak in the summer and early fall. The model provides a reasonable fit to the available data on mosquito abundance for the east side of Mauna Loa, Hawaii. The predictions of our model indicate the importance of abiotic conditions on mosquito dynamics and have important implications for the management of diseases transmitted by Cx. quinquefasciatus in Hawaii and elsewhere.

Venezuelan equine encephalitis (VEE) is a re-emerging, mosquito-borne viral disease with the potential to cause fatal encephalitis in both humans and equids. Recently, detection of endemic VEE caused by enzootic strains has escalated in Mexico, Peru, Bolivia, Colombia and Ecuador, emphasizing the importance of understanding the enzootic transmission cycle of the etiologic agent, VEE virus (VEEV). The majority of work examining the viral determinants of vector infection has been performed in the epizootic mosquito vector, Aedes (Ochlerotatus) taeniorhynchus. Based on the fundamental differences between the epizootic and enzootic cycles, we hypothesized that the virus-vector interaction of the enzootic cycle is fundamentally different from that of the epizootic model. We therefore examined the determinants for VEEV IE infection in the enzootic vector, Culex (Melanoconion) taeniopus, and determined the number and susceptibility of midgut epithelial cells initially infected and their distribution compared to the epizootic virus-vector interaction. Using chimeric viruses, we demonstrated that the determinants of infection for the enzootic vector are different than those observed for the epizootic vector. Similarly, we showed that, unlike A. taeniorhynchus infection with subtype IC VEEV, C. taeniopus does not have a limited subpopulation of midgut cells susceptible to subtype IE VEEV. These findings support the hypothesis that the enzootic VEEV relationship with C. taeniopus differs from the epizootic virus-vector interaction in that the determinants appear to be found in both the nonstructural and structural regions, and initial midgut infection is not limited to a small population of susceptible cells.

Larvicidal activity of ethyl acetate, butanol, and petroleum ether extracts of five species of Euphorbiaceae plants, Jatropha curcas, Pedilanthus tithymaloides, Phyllanthus amarus, Euphorbia hirta, and Euphorbia tirucalli, were tested against the early fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus (Say). The larval mortality was observed after 24 h of exposure. All extracts showed low larvicidal effects; however, the highest larval mortality was found in petroleum ether extract. The LC50 value of petroleum ether extracts of J. curcas, P. tithymaloides, P. amarus, E. hirta, and E. tirucalli were 8.79, 55.26, 90.92, 272.36, and 4.25 ppm, respectively, against A. aegypti and 11.34, 76.61, 113.40, 424.94, and 5.52 ppm, respectively, against C quinquefasciatus. Of the various ratios tested, the petroleum ether extracts of J. curcas and E. tirucalli were observed to be more efficient than the other plant extracts. It is, therefore, suggested that E. tirucalli can be applied as an ideal potential larvicide against A. aegypti and C. quinquefasciatus. This is an ideal ecofriendly approach for the control of the dengue vector, A. aegypti, and the lymphatic filariasis vector, C. quinquefasciatus.

We present data showing that structures serving as drains and catch basins for stormwater are important sources for production of the mosquito arbovirus vectors Aedes aegypti and Culex quinquefasciatus in Mérida City, México. We examined 1,761 stormwater drains – located in 45 different neighborhoods spread across the city – over dry and wet seasons from March 2012–March 2013. Of the examined stormwater drains, 262 (14.9%) held water at the time they were examined and 123 yielded mosquito immatures. In total, we collected 64,560 immatures representing nine species. The most commonly encountered species were Cx. quinquefasciatus (n=39,269) and Ae. aegypti (n=23,313). Ae. aegypti and Cx. quinquefasciatus were collected during all 11 months when we found water-filled stormwater drains, and both were found in stormwater drains located throughout Mérida City. We also present data for associations between structural characteristics of stormwater drains or water-related characteristics and the abundance of mosquito immatures. In conclusion, stormwater drains produce massive numbers of Ae. aegypti and Cx. quinquefasciatus across Mérida City, both in the wet and dry seasons, and represent non-residential development sites that should be strongly considered for inclusion in the local mosquito surveillance and control program. PMID:24582840

Background & objectives: Culex quinquefasciatus is the principal vector of lymphatic filariasis (LF). Application of alternative vector control methodologies are aimed at reduction of mosquito breeding sites and biting activity through the use of biological control methods. In the present study, functional response of aquatic Hemipteran backswimmer, Anisops sardea was assessed against Cx. quinquefasciatus larvae in laboratory bioassay. Methods: The functional respons of A. sardea was assessed against IIIrd instar larvae of Cx. quinquefasciatus. Respective handling times and coefficient of attack rates were determined by a non linear polynomeal regression equation. Results: The results of rates of predation in variable prey densities exhibited a ‘linear rise to plateau curve’, associated with ‘Type -II’ functional response. The logistic regression estimated a significant negative linear parameter (P1<0) which also supported the same observation. Associated ‘attack rates’ and ‘handling times’ were also calculated using the Holling Disc Equation. Interpretation & conclusions: The results of present experiments indicate that A. sardea can be used as a biocontrol agent against the larval forms of Cx. quinquefasciatus in temporarily available breeding places of mosquito with relatively clear water. However, a detailed field study has to be done to confirm these findings. PMID:25488451

Constituents in rosemary (Rosmarinus officinalis) have been shown to have larvicidal activity against invertebrates. In order to explore the properties of crude extract of rosemary further, we studied the chemical composition and its activity against dichlorodiphenyltrichloroethane (DDT)-susceptible, DDT-resistant, and field strains of Culex quinquefasciatus larvae. The major components of R. officinalis were found to be eucalyptol and camphor, with relative percentages of 10.93% and 5.51%, respectively. Minor constituents included limonene, (+)-4-carene, isoborneol, 1-methyl-4-(1-methylethylidene)-cyclohexene, and pinene. The median lethal concentration (LC50) values of the essential oil of R. officinalis against DDT-susceptible, DDT-resistant, and field strains of larvae of Cx. quinquefasciatus were 30.6, 26.4, and 38.3 mg/liter, respectively. The single median lethal dose (LD50) in Kunming mice was 4752 mg/kg. Essential oils from R. officinalis may, therefore, provide an effective natural plant product for use in mosquito prevention and control.

Like many insects, mosquitoes, rely on endosymbionts to grow and develop. These can be acquired from the environment. We used next generation 454 pyrosequencing to discern the whole-body microbiome of the mosquito species Culex quinquefasciatus in various larval stadia and following exposure to common pharmaceutical and personal care products (PPCPs) found in wastewater. PPCP treatments included environmentally-relevant concentrations; 1) a combination of common antibiotics, 2) a combination of mammalian hormones, 3) a mixture of the antibiotic and hormone treatments plus acetaminophen and caffeine and, 4) an untreated control. Within control groups, the predominant families of bacterial symbionts change with each larval instar despite consistent diets and rearing conditions. This trend was also seen in hormone treatments but not in the antibiotic or the mixture treatments. Richness and evenness were reduced in both antibiotic and mixture treatments, suggesting that antibiotics remove certain bacteria or inhibit them from increasing to proportions seen in the control treatment. Interestingly, the mixture treatments had greater richness and evenness compared to antibiotic alone treatments, possibly due to the other contaminants facilitating growth of different bacteria. These findings illuminate the complexity of the microbiome of C. quinquefasciatus and may have implications for more effective control strategies. PMID:26912375

The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1-A3) and four haplotypes (B1-B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1-AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1-AA4) by the COI gene and nine haplotypes (BB1-BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.

The mosquito-transmitted pathogen West Nile virus (WNV) is not yet present in the Galápagos Archipelago of Ecuador. However, concern exists for fragile endemic island fauna after population decreases in several North American bird species and pathology in certain reptiles. We examined WNV vector competency of a Galápagos strain of mosquito (Culex quinquefasciatus Say). Field specimens were tested for their capacity to transmit the WN02-1956 strain of WNV after incubation at 27°C or 30°C. Rates of infection, dissemination, and transmission all increased with days post-exposure to WNV, and the highest rates were observed at 28 days. Infection rates peaked at 59% and transmission rates peaked at 44% (of mosquitoes tested). Vector efficiency increased after day 14. Rates of infection but not of transmission were significantly influence by temperature. No vertical transmission was detectable. We demonstrate that Galápagos Cx. quinquefasciatus are competent WNV vectors, and therefore should be considered an animal and public health risk for the islands and controlled wherever possible. PMID:21896799

As an alternative to the use of commercially available, expanded-polystyrene beads, a study was made of the effectiveness of shredded, waste polystyrene (SWAP) for the control of Culex quinquefasciatus. The relevant physical properties of the SWAP were first investigated in the laboratory. Then, under field conditions in wet pit latrines, mosquito emergence rates were measured before and after application of the material. In the laboratory, when compared with the commercial product, the irregular shape of the SWAP particles greatly reduced their capacity to spread over the water surface and the interstitial air spaces also permitted respiration and development of mosquito larvae. Nevertheless, under field conditions, with careful application of the SWAP over the water surfaces in wet pit latrines, almost complete control of mosquito breeding was observed within a few days. This was sustained for 60 days, at which time observations were discontinued. SWAP appears to offer an effective, cheap and readily available alternative to the commercial product for the control of C. quinquefasciatus. The practicality of using it in community-based, mosquito-control programmes warrants further study.

Suction traps (30 cm diameter) were more effective for non-attractant sampling of flying adult Culex tarsalis Coquillett than were smaller CDC (5.5 cm diameter), Malaise or ramp traps. Comparative catch in suction traps operated in a variety of vegetation types indicated that females congregated along elevated ecotones and were significantly less abundant flying over low vegetation or under and over elevated vegetation. Most females taken at upland orchards or Tamarisk tree lines were unfed (97%, n = 5,278) and similar in reproductive condition to host-seeking females. Blood fed and gravid females and males were only abundant near emergence sites. Pyrocide 7396 (Pyrethrin 5%, PBO 25%) was applied at the label rate of 5 oz/min by truck mounted Pro-Mist ultra low volume (ULV) equipment and particle drift measured by bioassay. ULV particles dispersed well downwind over low vegetation, between citrus orchard rows, and under date orchard canopy, but did not penetrate citrus orchards or vineyards when rows were perpendicular to wind direction. Particles did move up and over vegetation contacting sentinel mosquitoes placed above the canopy. The congregation of adult mosquitoes at vegetative ecotones and within orchard vegetation may afford protection from ground applied ULV particles, negatively impacting control. These data may explain why repeated applications often fail to interrupt encephalitis virus transmission.

Mark-release-recapture experiments were conducted to determine the length of the gonotrophic cycle and rate of survivorship of Culex quinquefasciatus Say in Monterrey, northeastern Mexico. A total of 2,352 field-caught Cx. quinquefasciatus females were marked and released at 8-12 h postemergence in 2 field trials. Sticky ovitraps were used to recapture marked gravid females. One hundred and ten (4.6%) marked females were recaptured during a 12-day sampling period. Recapture rates for the 2 individual trials were 6.4% and 3.5%. The length of the gonotrophic cycle, calculated as the average time between the initial blood meal and the time of recapture of gravid females, was 2-3 days. The first blood-fed mosquitoes were recaptured on the 2nd day postrelease. Gravid egg-laying females were most commonly recaptured at 2-3 days postfeeding. Daily survival estimates for the 2 release dates were of 0.871 and 0.883, respectively.

Resistance of mosquitoes to insecticides is a growing concern in India. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programs mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. In the present study, we evaluate the permethrin resistance and cross-resistance pattern of several insecticides in Culex quinquefasciatus mosquitoes. After 10 generation of selection with permethrin, the LC50 value for both larvae and adult Cx. quinquefasciatus was increased by 17.3- and 17.1-folds compared with susceptible strain. Detoxification enzyme profiles and native PAGE electrophoresis of esterase isoenzyme further revealed that esterase and CytP450 may be involved in permethrin resistance (PerRes) strain compared with susceptible strain. In addition to cross-resistance, study revealed that high resistance to cypermethrin (RR = 6.3, 8.8-folds). This study provided important information for understanding permethrin resistance and facilitating a better strategy for the management of resistance. These studies conclude that a strong foundation for further study of permethrin resistance mechanisms observed in Cx. quinquefasciatus mosquitoes.

In temperate regions, seasonal epidemics of many mosquito-borne viruses are triggered when mosquito populations shift from feeding on avian to mammalian hosts. We investigated effects of temperature on the timing of bird-to-mammal shifts using an 8 year dataset of blood-meals from a mosquito (Culex erraticus) in Alabama, USA. As expected, Cx. erraticus shifted from avian to mammalian hosts each year. The timing of the shift, however, varied considerably among years. Harshness of the preceding winter (chill accumulation) explained 93 per cent of the variation in the timing of bird-to-mammal shifts, with shifts occurring later in years following harsher winters. We hypothesize that winter temperatures drive the timing of bird-to-mammal shifts through effects on host reproductive phenology. Because mosquitoes target birds during the nesting season, and bird nesting occurs later in years following colder winters, later nesting dates result in a concomitant delay in the timing of bird-to-mammal host shifts. Global increases in winter temperatures could cause significant changes in the timing of seasonal host shifts by mosquitoes, with prolonged periods of epidemic transmission of mosquito-borne diseases. PMID:22399787

Studies were conducted to determine the host-feeding preference of Culex quinquefasciatus Say (Diptera: Culicidae) in relation to the availability of human and domestic animals in the city of Merida, Yucatan State, Mexico. Mosquitoes were collected in the backyards of houses using resting wooden boxes. Collections were made five times per week from January to December 2005. DNA was extracted from engorged females and tested by PCR using universal avian- and mammalian-specific primers. DNA extracted from avian-derived blood was further analyzed by PCR using primers that differentiate among the birds of three avian orders: Passeriformes, Columbiformes and Galliformes. PCR products obtained from mammalian-derived blood were subjected to restriction enzyme digestion to differentiate between human-, dog-, cat-, pig-, and horse-derived blood meals. Overall, 82% of engorged mosquitoes had fed on birds, and 18% had fed on mammals. The most frequent vertebrate hosts were Galliformes (47.1%), Passeriformes (23.8%), Columbiformes (11.2%) birds, and dogs (8.8%). The overall human blood index was 6.7%. The overall forage ratio for humans was 0.1, indicating that humans were not a preferred host for Cx. quinquefasciatus in Merida.

The present study was carried out to establish the larvicidal properties of crude extracts of three plants, viz. Carica papaya, Murraya paniculata and Cleistanthus collinus against Culex quinquefasciatus as target species. The relative efficacy of the plant extracts in vector control was as follows: C. papaya seed extract > M. paniculata fruit extract > M. paniculata leaf extract > C. collinus leaf extract. Result of log-probit analysis (at 95% confidence level) revealed that LC(50) and LC(95) values gradually decreased with the exposure periods similar to the result in bioassay experiment for each of the plant extract. The results of preliminary qualitative phytochemical analysis of all the plants revealed the presence of many bioactive principles such as steroids, alkaloids, terpenes, saponins, etc. that may be responsible for their biocontrol potentiality. The appropriate lethal concentrations at 24 h for each of the extracts were also studied on non-target organisms such as Diplonychus annulatum and Chironomus circumdatus, and no change in the swimming behaviour and survivality was noticed, which indicates that all the extracts are safe for the aquatic ecosystem.

Elevated esterase Estbeta1 was purified from larvae of newly isolated strains of the mosquito Culex quinquefasciatus from Colombia (COL) and Trinidad (TRI) with resistance to organophosphate (OP) insecticides. Insecticide interactions were compared with those of elevated Estbeta1(2) from the OP-resistant Habana strain and the non-elevated Estbeta1(3) from the susceptible PelSS strain. On the basis of insecticide binding efficiency, all elevated Estbeta1 esterases were readily distinguishable. Differences between the EcoRI restriction fragment patterns of the amplified estbeta1 gene in COL and TRI strains compared with each other, and between amplified estbeta1(1), estbeta1(2) and the non-amplified estbeta1(3), suggest differences in their nucleotide sequence. Considering their variable insecticide binding efficiencies, these genetic differences would imply that, in contrast to estalpha2 and estbeta2, amplification of estbeta1 has occurred several times independently. Generally, the elevated Estbeta1s were more reactive with insecticides than the non-elevated Estbeta1(3). This supports the hypothesis that the elevated esterase-based mechanism confers resistance through amplification of alleles coding for esterases which have a greater specificity for the insecticides they sequester than the esterases coded by their non-amplified counterparts.

The impact of nutritional variables on the development of host-seeking and biting behaviours after emergence by female Culex nigripalpus mosquitoes were studied using air-flow olfactometer and close-range biting assays, respectively. Unfed females failed to develop resting stage ovarian follicles. When offered a bird host in the absence of competing stimuli, sugar-fed mosquitoes were significantly more responsive in both host-seeking and biting than unfed controls. In a choice olfactometer assay using nectar odours (honey scented with artificial apple-blossom oil) versus host odours (a bird), unfed females preferred honey over bird odours except when honey odour was weak. After sucrose feeding, females switched from honey to bird preference. This change in behaviour was accompanied by significant accumulation of lipid and by follicular growth to the resting stage. Elevation of host responsiveness after sugar feeding was reversible; starvation ultimately resulted in females preferring honey over bird odours. When the larval diet was restricted by crowding, the wing-length and total lipid of resultant adult females were reduced. Although differences were subtle, unfed bird-responding females tended to have longer wings and more lipid than their honey-responding counterparts.

A set of 67 novel LTR-retrotransposon has been identified by in silico analyses of the Culex quinquefasciatus genome using the LTR_STRUC program. The phylogenetic analysis shows that 29 novel and putatively functional LTR-retrotransposons detected belong to the Ty3/gypsy group. Our results demonstrate that, by considering only families containing potentially autonomous LTR-retrotransposons, they account for about 1% of the genome of C. quinquefasciatus. In previous studies it has been estimated that 29% of the genome of C. quinquefasciatus is occupied by mobile genetic elements. The potential role of retrotransposon insertions strictly associated with host genes is described and discussed along with the possible origin of a retrotransposon with peculiar Primer Binding Site region. Finally, we report the presence of a group of 38 retrotransposons, carrying tandem repeated sequences but lacking coding potential, and apparently lacking “master copy” elements from which they could have originated. The features of the repetitive sequences found in these non-autonomous LTR retrotransposons are described, and their possible role discussed. These results integrate the existing data on the genomics of an important virus-borne disease vector. PMID:22383973

Twenty local isolates of entomopathogenic fungi were determined for control of the larvae and adults of Culex quinquefasciatus. In a laboratory experiment, a Penicillium sp. CM-010 caused 100% mortality of third-instar larvae within 2 h using a conidial suspension of 1 × 10⁶ conidia ml⁻¹. Its LC₅₀ was 3 × 10⁵ conidia ml⁻¹, and the lethal time (LT₅₀) was 1.06 h. Cloning and sequencing of its internal transcribed spacer region indicated that this Penicillium species is Penicillium citrinum (100% identity in 434 bp). Mortality of the adult was highest with Aspergillus flavus CM-011 followed with Metarhizium anisopliae CKM-048 from 1 × 10⁹ conidia ml⁻¹. P. citrinum CM-010 at 1 × 10⁶ conidia ml⁻¹ killed 100% larvae within 2 h while Bacillus thuringiensis var. israelensis at 5 ITU ml⁻¹ required 24 h. This P. citrinum CM-010 also greatly reduced survival of C. quinquefasciatus larvae in an unreplicated field test. Light and transmission electron micrographs showed that the fungal conidia were ingested by the larvae and deposited in the gut. The metabolite patulin was produced by P. citrinum CM-010 instead of citrinin.

The Culex quinquefaciatus Say, commonly known as the southern house mosquito, is well known for biting nuisance and vectoring of some fatal diseases. Synthetic chemicals have been relied upon as the major control measure to control mosquitoes. Therefore, we have evaluated 21 insecticides belonging to different chemical classes for their toxicity to C. quinquefaciatus females. Chlorfenapyr was the most toxic adulticide among all the tested insecticides. Among pyrethroids, deltamethrin was the least toxic adulticide, and all other have same toxicity. In case of organophosphates, the chlorpyrifos was the most toxic insecticide. Neonicotinoids such as acetamiprid, nitenpyram, and clothianidin have similar toxicity based on overlapping of 95 % confidence intervals (CI) and were more toxic when compared with the imidacloprid. The spinetoram was more toxic as compared with the spinosad (based on non-overlapping 95 % Cl). In case of ketoenoles, spirotetrament was more toxic as compared with the spiromesifen. Emamectin benzoate was the most toxic insecticide when compared with fipronil and indoxacarb. We also have tested four insect growth regulators (IGRS) including lufenuron, methoxyfenozide, pyriproxyfen, and cyromazine as larvicides. The lufenuron and pyriproxyfen have similar toxicity based upon their overlapping 95 % CI and were more toxic as compared with the methoxyfenozide and cyromazine. The methoxyfenozide was the moderately toxic among all the tested IGRS, and cyromazine was the least toxic among all the tested IGRS. These results will prove helpful in effectuating an effective integrated vector management program for C. quinquefaciatus.

The diel biting periodicity of the arbovirus vector Culex (Melanoconion) caudelli Dyar and Knab (Diptera; Culicidae) in the Aripo-Wallerfield forest, Trinidad, was studied by collecting mosquitoes attracted to mouse-baited traps at 2-hourly intervals during eight 24 h periods. Biting females of Cx caudelli were collected during all night-time hours, 18.00-06.00 hours, with an overall unimodal pattern, i.e. one well-defined peak between 22.00 and 04.00 hours. This contrasts with previous reports that Cx caudelli is diurnally active. During the four moon phases, the period of peak biting activity varied from 22.00 to 24.00 hours at fullmoon, from 22.00 to 04.00 hours at first quarter, and from 24.00 to 02.00 hours at new moon and last quarter phases. The number of mosquitoes collected varied significantly with moon phases, the highest (152/24 h) being collected in the first lunar quarter and the lowest (71/24 h) in the last lunar quarter. Cloud cover and rainfall had no significant effect on the diel biting periodicity of Cx caudelli.

Despite their importance as vectors of human and livestock diseases, relatively little is known about innate antiviral immune pathways in mosquitoes and other insects. Previous work has shown that Culex Vago (CxVago), which is induced and secreted from West Nile virus (WNV)-infected mosquito cells, acts as a functional homolog of interferon, by activating Jak-STAT pathway and limiting virus replication in neighbouring cells. Here we describe the Dicer-2-dependent pathway leading to WNV-induced CxVago activation. Using a luciferase reporter assay, we show that a NF-κB-like binding site in CxVago promoter region is conserved in mosquito species and is responsible for induction of CxVago expression following WNV infection. Using dsRNA-based gene knockdown, we show that the NF-κB ortholog, Rel2, plays significant role in the signaling pathway that activates CxVago in mosquito cells in vitro and in vivo. Using similar approaches, we also show that TRAF, but not TRAF-3, is involved in activation of Rel2 after viral infection. Overall the study shows that a conserved signaling pathway, which is similar to mammalian interferon activation pathway, is responsible for the induction and antiviral activity of CxVago.

Two preparations of Bacillus sphaericus 2362 were studied for their biological activity, delayed mortality and the induction of morphogenetic aberrations in larvae, pupae and adults of Culex quinquefasciatus. Longevity and fecundity of adult mosquitoes were also assessed. A dosage response line for B. sphaericus was established against 4th-instar larvae and sublethal concentrations (48 h LC50 and lower) were used against these larvae. Sublethal concentrations of B. sphaericus induced delayed mortality in larvae, pupae and adults. The magnitude of mortality increased in succeeding cohorts and developmental stages resulting from the surviving larvae. Only 10 and 25% overall emergence of viable adults occurred in the sublethal treatments (LC25) of 2 B. sphaericus preparations. The range of successful adult emergence was over 94% in the controls. A wide range of external morphogenetic aberrations in dead larvae, pupae and adults were noted. These aberrations and gross morphological features were quite similar to those reported for certain insect growth regulators. Sublethal concentrations had no marked effect on longevity of adults, egg deposition and hatch.

The mosquito-transmitted pathogen West Nile virus (WNV) is not yet present in the Galápagos Archipelago of Ecuador. However, concern exists for fragile endemic island fauna after population decreases in several North American bird species and pathology in certain reptiles. We examined WNV vector competency of a Galápagos strain of mosquito (Culex quinquefasciatus Say). Field specimens were tested for their capacity to transmit the WN02-1956 strain of WNV after incubation at 27°C or 30°C. Rates of infection, dissemination, and transmission all increased with days post-exposure to WNV, and the highest rates were observed at 28 days. Infection rates peaked at 59% and transmission rates peaked at 44% (of mosquitoes tested). Vector efficiency increased after day 14. Rates of infection but not of transmission were significantly influence by temperature. No vertical transmission was detectable. We demonstrate that Galápagos Cx. quinquefasciatus are competent WNV vectors, and therefore should be considered an animal and public health risk for the islands and controlled wherever possible.

Studies were conducted to determine the host-feeding preference of Culex quinquefasciatus Say (Diptera: Culicidae) in relation to the availability of human and domestic animals in the city of Merida, Yucatan State, Mexico. Mosquitoes were collected in the backyards of houses using resting wooden boxes. Collections were made five times per week from January to December 2005. DNA was extracted from engorged females and tested by PCR using universal avian- and mammalian-specific primers. DNA extracted from avian-derived blood was further analyzed by PCR using primers that differentiate among the birds of three avian orders: Passeriformes, Columbiformes and Galliformes. PCR products obtained from mammalian-derived blood were subjected to restriction enzyme digestion to differentiate between human-, dog-, cat-, pig-, and horse-derived blood meals. Overall, 82% of engorged mosquitoes had fed on birds, and 18% had fed on mammals. The most frequent vertebrate hosts were Galliformes (47.1%), Passeriformes (23.8%), Columbiformes (11.2%) birds, and dogs (8.8%). The overall human blood index was 6.7%. The overall forage ratio for humans was 0.1, indicating that humans were not a preferred host for Cx. quinquefasciatus in Merida. PMID:20578953

Culex salinarius is considered one of the most likely bridge vectors involved in the human transmission cycle of West Nile virus (WNV) and eastern equine encephalomyelitis virus (EEEV) in the northeastern USA. The larval habitats of this species in the coastal region of New York State are currently poorly known. Between 2005 and 2007, a larval survey was carried out to identify and characterize possible larval habitats in Suffolk County, encompassing natural and man-made freshwater wetlands, artificial containers, and salt marshes. Only relatively undisturbed salt marsh yielded Cx. salinarius larvae in considerable numbers from several sites over a period of 2 years. The immature stages of this species were found associated with Spartina patens and S. alterniflora of the upper marsh at salinities ranging from 4.3 to 18.8 parts per thousand. Both heavily impacted and relatively undisturbed salt marshes produced several hundreds of adult Cx. salinarius per Centers for Disease Control and Prevention (CDC) light trap per night, an order of magnitude higher than CDC light traps deployed at upland sites. The ability of Cx. salinarius to use both heavily impacted and relatively undisturbed salt marshes for reproduction has significant repercussions for marsh restoration and vector control practices.

Simultaneous recording of electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) has shown a number of advantages that make this multimodal technique superior to fMRI alone. The feasibility of recording EEG at ultra-high static magnetic field up to 9.4 T was recently demonstrated and promises to be implemented soon in fMRI studies at ultra high magnetic fields. Recording visual evoked potentials are expected to be amongst the most simple for simultaneous EEG/fMRI at ultra-high magnetic field due to the easy assessment of the visual cortex. Auditory evoked P300 measurements are of interest since it is believed that they represent the earliest stage of cognitive processing. In this study, we investigate the feasibility of recording visual evoked potentials and auditory evoked P300 in a 9.4 T static magnetic field. For this purpose, EEG data were recorded from 26 healthy volunteers inside a 9.4 T MR scanner using a 32-channel MR compatible EEG system. Visual stimulation and auditory oddball paradigm were presented in order to elicit evoked related potentials (ERP). Recordings made outside the scanner were performed using the same stimuli and EEG system for comparison purposes. We were able to retrieve visual P100 and auditory P300 evoked potentials at 9.4 T static magnetic field after correction of the ballistocardiogram artefact using independent component analysis. The latencies of the ERPs recorded at 9.4 T were not different from those recorded at 0 T. The amplitudes of ERPs were higher at 9.4 T when compared to recordings at 0 T. Nevertheless, it seems that the increased amplitudes of the ERPs are due to the effect of the ultra-high field on the EEG recording system rather than alteration in the intrinsic processes that generate the electrophysiological responses.

This study evaluated whether listeners can distinguish human brainstem auditory evoked responses elicited by acoustic clicks from control waveforms obtained with no acoustic stimulus when the waveforms are presented auditorily. Detection performance for stimuli presented visually was slightly, but consistently, superior to that which occurred for…

Results of somatosensory evoked potential (SEP) and brainstem auditory evoked potential (BAEP) examinations performed early in the clinical course of patients with acute cerebrovascular disease correlate statistically significantly with outcome regardless of type and localization of the primary lesion. The prognostic value of serial examinations of SEP and BAEP has not been studied yet. The authors examined a group of 215 patients suffering from acute stroke requiring neurocritical care composed of 75 supratentorial and 36 infratentorial ischemic strokes, 58 supratentorial and 18 infratentorial hemorrhages, and 28 aneurysmatic subarachnoid hemorrhages prospectively using spinal and cortical SEP and BAEP according to routine procedures on admission as well as after 1 and 2 weeks. The findings were correlated to outcome at 4 weeks. Statistical assessment was performed using standard methods of contingency analysis. In all groups, SEP findings were significantly correlated with outcome at initial and all subsequent examinations, similar correlations were also found for BAEP. However, after partialling out the prognostic information gained from the initial examination of SEP and BAEP, the follow-up examinations rendered only a marginal increase in prognostic information. Therefore, the initial examination of evoked potentials supplies valuable prognostic information, however, serial examinations of evoked potentials during the first weeks of disease improve the prognostic information only marginally.

Early diagnosis of spondylodiscitis is a condition of efficient conservative treatment. Somatosensory-evoked potentials with clinical examination results are used in assessing the diagnosis, as well as in monitoring the course of disease and healing. MRI clearly shows the inflammatory process, healing and scars. We report a 46-year-old woman suffering from non-specific interscapular pains. The evoked somatosensory potentials of the tibial nerveshow potential conductivity being slowed down through the thoracic spine, which is clearly evident from the prolonged latency and the decreased amplitude of the evoked response. The performed thoracic spine MRI shows spondylodiscitis at the Thl0-11 level. The subject is a nurse administering BCG therapy at a urology clinic, due to the fact of which this was deemed to have been a case of tuberculous spondylodiscitis. Due to the possibility of scattering the causative agent by needle, the biopsy was given up and antituberculous therapy was administered ex juvantibus. The disease was followed up by clinical examinations, somatosensory-evoked potentials and MRI up to fully successful and final recovery from spondylodiscitis. The above examinations are of great help in diagnosing the tuberculous spondylodiscitis and monitoring the recovery (Fig. 6, Ref. 16).

In this paper we have studied cortical dynamics as assessed using graphical methods during deep anaesthesia. Graphical analysis was carried out by autocorrelation functions and return maps with different lags. During moderate and deep anaesthesia, the electroencephalogram (EEG) shows a burst suppression pattern, consisting of abruptly-occurring high amplitude bursts alternating with periods of relative silence. Deep anaesthesia with burst suppression pattern provides a simple model of brain activity when the noise that is usually present in a subject who is awake is suppressed. During anaesthesia-induced EEG suppression, the brain reacts to different external stimuli with bursts. In respect to sensory processing during anaesthesia, it is interesting to know whether these bursts have different dynamics depending on the stimuli used. We have used graphical analysis to reveal the possible differences in bursts evoked by different stimuli. Externally evoked bursts were induced by auditory, electric and visual stimuli. The EEG studied in this paper consists of 25 bursts from one subject. We have estimated the autocorrelation function for each burst and used the formation gained from such autocorrelation coefficients as the grounds for determining different lags for return maps. The graphical methods used revealed differences in dynamics and topology of bursts as evoked by different stimuli. Spontaneous bursts clearly had different dynamics from evoked burst; which could not be seen directly from the raw EEG data. This study suggests that graphical analysis is a useful tool to obtain information about the dynamics of neuronal processes behind cortical responses during deep anaesthesia.

The present study investigated the effects of differential negative reinforcement of other behavior (DNRO) on problem behavior evoked by music in a 7-year-old child with pervasive developmental disorder. Following an auditory stimulus assessment, DNRO was used to reduce problem behavior to near-zero levels. Results are discussed in terms of…

In some cochlear implant users, success is not achieved in spite of optimal clinical factors (including age at implantation, duration of rehabilitation and post-implant hearing level), which may be attributed to disorders at higher levels of the auditory pathway. We used cortical auditory evoked potentials to investigate the ability to perceive…

In order to optimize procedure for the assessment of evoked potentials and to provide visualization of the flow of action potentials along the motor systems, we introduced array electrodes for stimulation and recording and developed software for the analysis of the recordings. The system uses a stimulator connected to an electrode array for the generation of evoked potentials, an electrode array connected to the amplifier, A/D converter and computer for the recording of evoked potentials, and a dedicated software application. The method has been tested for the assessment of the H-reflex on the triceps surae muscle in six healthy humans. The electrode array with 16 pads was positioned over the posterior aspect of the thigh, while the recording electrode array with 16 pads was positioned over the triceps surae muscle. The stimulator activated all the pads of the stimulation electrode array asynchronously, while the signals were recorded continuously at all the recording sites. The results are topography maps (spatial distribution of evoked potentials) and matrices (spatial visualization of nerve excitability). The software allows the automatic selection of the lowest stimulation intensity to achieve maximal H-reflex amplitude and selection of the recording/stimulation pads according to predefined criteria. The analysis of results shows that the method provides rich information compared with the conventional recording of the H-reflex with regard the spatial distribution.

Two alternative approaches for studying short-latency click-evoked otoacoustic emission (OAE) in normal-hearing subjects were employed. Growth of a click-evoked "ear canal response" with stimulus increase became progressively more non-linear and saturated when the latency of the analyzed segment of response increased. This relation between latency and shape of the response input/output function was observed even after linear component cancellation, indicating that it could be an intrinsic property of OAE. Hence, the existence of an essentially linear short-latency OAE component which is probably eliminated by commonly used artifact cancellation technique is suggested. Taking into account the fact that transient evoked otoacoustic emission (TEOAE) may be completely suppressed by simultaneously presented noise, a "true" artifact cancellation was performed by subtracting the ear canal response in the presence of a masker from the conventional click-evoked OAE recording. An additional TEOAE component with a latency of 2.5-5 ms was found. Its growth with stimulus intensity was indeed more linear than that of later components. However, latency and frequency of this TEOAE component, being specific for each subject, can hardly be explained by both a commonly assumed latency-frequency relationship of TEOAE and a generally used estimation of TEOAE latency as the sum of the forward and backward traveling wave propagation times.

Previous findings have shown that individuals with autism spectrum disorder (ASD) evince greater intra-individual variability (IIV) in their sensory-evoked fMRI responses compared to typical control participants. We explore the robustness of this finding with a new sample of high-functioning adults with autism. Participants were presented with…

The authors examined electroencephalographic (EEG) oscillatory responses to two violent events, the player character wounding and killing an opponent character with a gun, in the digital game James Bond 007: NightFire. EEG was recorded from 25 (16 male) right-handed healthy young adults. EEG data were segmented into one 1-s baseline epoch before each event and two 1-s epochs after event onset. Power estimates (microV(2)) were derived with the fast Fourier transform (FFT) for each artefact free event. Both of the studied events evoked increased occipital theta (4-6Hz) responses as compared to the pre-event baseline. The wounding event evoked also increased occipital high theta (6-8Hz) response and the killing event evoked low alpha (8-10Hz) asymmetry over the central electrodes, both relative to the pre-event baseline. The results are discussed in light of facial electromyographic and electrodermal activity responses evoked by these same events, and it is suggested that the reported EEG responses may be attributable to affective processes related to these violent game events.

To investigate whether individuals' ongoing neuronal activity at resting state can affect their response to brain stimulation, fMRI BOLD signals were imaged from the human visual cortex of fifteen healthy subjects in the absence and presence of visual stimulation. It was found that the temporal correlation strength but not amplitude of baseline BOLD signal fluctuations acquired under the eyes-fixed condition is positively correlated with the amplitude of stimulus-evoked BOLD responses across subjects. Moreover, the spatiotemporal correlations of baseline BOLD signals imply a coherent network covering the visual system, which is topographically indistinguishable from the "resting-state visual network" observed under the eyes-closed condition. The overall findings suggest that the synchronization of ongoing brain activity plays an important role in determining stimulus-evoked brain activity even at an early stage of the sensory system. The tight relationship between baseline BOLD correlation and stimulus-evoked BOLD amplitude provides an essential basis for understanding and interpreting the large inter-subject BOLD variability commonly observed in numerous fMRI studies and potentially for improving group fMRI analysis. This study highlights the importance to integrate the information from both resting-state coherent networks and task-evoked neural responses for a better understanding of how the brain functions.

Auditory brainstem evoked responses (ABRs) were compared in 15 newborns with Down syndrome and 15 sex-, age-, and weight-matched control newborns. Participants had normal ABRs based upon values specific to 32- to 42-weeks postconceptional age. Although Wave III and Wave V component latencies and the Wave I-III interpeak latency (IPL) were shorter…

Auditory evoked potentials are tools widely used to assess auditory cortex functions in clinical context. However, in cochlear implant users, electrophysiological measures are challenging due to implant-created artefacts in the EEG. Here, we used independent component analysis to reduce cochlear implant-related artefacts in event-related EEGs of…

The ‘thermal grill illusion’ (TGI) is a unique cutaneous sensation of unpleasantness, induced through the application of interlacing warm and cool stimuli. While previous studies have investigated optimal parameters and subject characteristics to evoke the illusion, our aim was to examine the modulating effect as a conditioning stimulus. A total of 28 healthy control individuals underwent three testing sessions on separate days. Briefly, 15 contact heat stimuli were delivered to the right hand dorsum, while the left palmar side of the hand was being conditioned with either neutral (32 °C), cool (20 °C), warm (40 °C), or TGI (20/40 °C). Rating of perception (numeric rating scale: 0–10) and evoked potentials (i.e., N1 and N2P2 potentials) to noxious contact heat stimuli were assessed. While cool and warm conditioning decreased cortical responses to noxious heat, TGI conditioning increased evoked potential amplitude (N1 and N2P2). In line with other modalities of unpleasant conditioning (e.g., sound, visual, and olfactory stimulation), cortical and possibly sub-cortical modulation may underlie the facilitation of contact heat evoked potentials. PMID:28079118

Flash-evoked responses can be recorded from the fetus in utero. However, a standard analysis approach based on orthogonal projection (OP) to attenuate maternal and fetal cardiac signals leads to a spatial redistribution of the signal. This effect prevents the correlation of source location with a known fetal head location in some cases and the signal-to-noise ratio (SNR) is sometimes limited such that the response latency is difficult to determine. We used a modified beamformer model search analysis to avoid the redistribution shortcoming and to improve the SNR. We included a statistical test for residual interference in the average and quantified significance of the evoked response with a bootstrap method. Selected source locations compared favorably to fetal head locations estimated from ultrasound exams. The evoked response time course was found to have a significant post-trigger peak with a latency between about 180 and 770 ms in more than 90% of the subject measurements. These results confirm that the combined application of a beamformer model search and bootstrap significance test provides a validation of the flash-evoked response observed in OP processed fetal MEG channels.

The locations of cortical activity evoked by visual stimuli presented at different positions in the visual field are deduced from the scalp topography of visually evoked potentials in humans. To accomplish this, the Laplacian evoked potential is measured using a multi-electrode array. It is shown that the Laplacian response has the following useful attributes for this purpose. It is reference-free. Its spatial resolution is approximately 2 cm referred to the surface of the cortex. Its spatial sensitivity characteristic is that of a spatial band-pass filter. It is relatively insensitive to source--sink configurations that are oriented tangentially to the surface of the scalp. Only modest assumptions about the source--sink configuration are required to obtain a unique inversion of the scalp topography. Stimuli consisting of checkerboard-filled octant or annular octant segments are presented as appearance-disappearance pulses at sixteen different positions in the visual field in randomized order. The locations of evoked cortical activity in the occipital, parietal and temporal lobes are represented on a Mercator projection map for each octant or octant segment stimulated. Lower hemifield stimuli activate cortex which lies mainly on the convexity of the occipital lobe contralateral to the side of stimulus presentation in the visual field. The more peripheral the stimulus is in the visual field, the more rostral is the location of the active cortex. The rostral-to-caudal location of the evoked activity varies from subject to subject by as much as 3 cm on the surface of the occipital cortex. Furthermore, in any single subject there is a substantial amount of hemispheric asymmetry. Upper hemifield stimuli activate cortex that lies on the extreme caudal pole of the occipital lobe. This activity is relatively weak, and in some subjects it is almost unmeasurable. It is suggested that the representation of the upper hemifield in the cortex lies mostly on the inferior and

Arsenic contamination of global water supplies has come to the forefront in policy decisions in recent decades. However, the effects of arsenic on lower trophic levels of insects inhabiting contaminated ecosystems are not well understood. One approach to document both acute and sublethal effects of toxicants like arsenic is to assay them in combination with microbial pathogens to evaluate shifts in survival curves of the test organisms. Larvae of Culex quinquefasciatus and Culex tarsalis were reared in water containing 0 or 1 000 μg/L of arsenate or arsenite. Fourth instars were then exposed to a range of doses of Bacillus thuringiensis subsp. israelensis (Bti) or Lysinibacillus sphaericus (Ls), with shifts in lethal concentrations determined. Arsenic accumulation in 4th instars was also quantified, and a relative growth index (RGI) calculated for the treatments and compared to controls. Larvae of both species accumulated between 4 447 ± 169 ng As/g and 6 983 ± 367 ng As/g, though RGI values indicated accumulation did not affect growth and development. In all cases, the LC50 's and LC90 's of Cx. quinquefasciatus exposed jointly with arsenic and Bti/Ls were higher than Cx. tarsalis. Cx. tarsalis reared in arsenite showed a significant reduction in their Bti LC90 values compared to the control, indicating a sublethal effect of Bti. When exposed jointly with Ls, arsenite was more toxic than arsenate in Cx. tarsalis. Overall, these results indicate tolerance of these Culex species to arsenic exposures, and why this may occur is discussed.

Mosquitoes are responsible for spread of many diseases than any other group of arthropods. Diseases such as malaria, filariasis, dengue hemorrhagic fever (DHF), and chikunguinya are real threat to mankind. In the present study, ethanolic extracts of leaves of Pongamia pinnata, Coleus forskohlii, and Datura stramonium were evaluated for oviposition deterrent activity against Aedes aegypti and Culex quinquefasciatus. The oviposition deterrent tests of ethanolic extract of Pongamia pinnata, Coleus forskohlii, and Datura stramonium leaves reduced egg laying by 97.62%, 77.3%, 100% against Aedes aegypti and 59.10%, 39.22%, 82% against Culex quinquefasciatus at higher concentration (0.1%).

The superficial layers of the frog optic tectum receive a projection from the contralateral eye that forms a point-to-point map of the visual field. The monocular part of the visual field of the contralateral eye is represented in the caudolateral region of the tectum while the binocular part of the visual field is represented in the rostromedial tectum. Within the representation of the binocular field (rostromedial tectum), the maps of visual space from each eye are aligned. The tectal representation of the binocular visual field of the ipsilateral eye is mediated through a crossed projection from the midbrain nucleus isthmi. This isthmotectal projection also terminates in the caudolateral region of the optic tectum, yet there has been no indication that it forms a functional connection. By extracellular recording in intermediate layer 7 of the caudolateral tectum, we have discovered electrical activity driven by visual stimulation in the monocular visual field of the ipsilateral eye. The units driven from the ipsilateral eye burst upon initial presentation of the stimulus. At individual layer 7 recording sites in the caudolateral tectum, the multiunit receptive field evoked from the ipsilateral eye is located at the mirror image spatial location to the multiunit receptive field driven by the contralateral eye. Thus, as revealed electrophysiologically, there are superimposed topographic maps of the monocular visual fields in the caudolateral tectum. The ipsilateral eye monocular visual field representation can be abolished by electrolytic ablation of contralateral nucleus isthmi.

A successful experimental infection of Culex quinquefasciatus with Hepatozoon sp. infecting Cerastes cerastes cerastes viper was carried out under laboratory conditions of 24 +/- 3 degrees C and 60-70% R. H. The period monitored for complete sporogonic cycle was 21 days. The effect of high parasitimic blood meal was nonsignificant (P>0.05) on preoviposition period and hatchability. Meanwhile a highly significant reduction was observed in oviposition rate, number of deposited eggs, number of hatched larvae and longevity (P<0.01). On the contrary moderate infection with Hepatozoon revealed a great significant increase in fecundity (P<0.01) and a nonsignificant decrease in longevity (P>0.05)

Introduced mosquito-borne pathogens avian malaria (Plasmodium relictum Grassi and Feletti) and avian pox virus (Avipoxvirus) have been implicated in the past extinctions and declines of Hawaiian avifauna and remain significant obstacles to the recovery and restoration of endemic Hawaiian birds. Effective management of avian disease will require extensive mosquito control efforts that are guided by the local ecology of the vector Culex quinquefasciatus Say (Diptera: Culicidae). During October and November 1997 and September through November 1998 five mark-release-recapture experiments with laboratory-reared Cx. quinquefasciatus were conducted in a native rain forest on Hawaii Island. Of the overall 66,047 fluorescent dye-marked and released females, 1,192 (1.8%) were recaptured in 43-52 CO2-baited traps operated for 10-12-d trapping periods. Recaptured mosquitoes were trapped in all directions and at distances up to 3 km from the release site. The cumulative mean distance traveled (MDTs) over the trapping period ranged from a high of 1.89 km after 11 d (September 1998) to a low of 0.81 km after 11 d (November 1998). Released mosquitoes moved predominately in a downwind direction and they seemed to use forestry roads as dispersal corridors. Applying an estimated MDT of 1.6 km to a geographical information system-generated map of the Hakalau Forest National Wildlife Refuge clearly demonstrated that the effective refuge area could be reduced 60% by mosquitoes infiltrating into managed refuge lands. These findings should have significant implications for the design of future refuges and development of effective mosquito-borne avian disease control strategies.

We present a population model to understand the effects of temperature and rainfall on the population dynamics of the southern house mosquito, Culex quinquefasciatus Say, along an elevational gradient in Hawaii. We use a novel approach to model the effects of temperature on population growth by dynamically incorporating developmental rate into the transition matrix, by using physiological ages of immatures instead of chronological age or stages. We also model the effects of rainfall on survival of immatures as the cumulative number of days below a certain rain threshold. Finally, we incorporate density dependence into the model as competition between immatures within breeding sites. Our model predicts the upper altitudinal distributions of Cx. quinquefasciatus on the Big Island of Hawaii for self-sustaining mosquito and migrating summer sink populations at 1,475 and 1,715 m above sea level, respectively. Our model predicts that mosquitoes at lower elevations can grow under a broader range of rainfall parameters than middle and high elevation populations. Density dependence in conjunction with the seasonal forcing imposed by temperature and rain creates cycles in the dynamics of the population that peak in the summer and early fall. The model provides a reasonable fit to the available data on mosquito abundance for the east side of Mauna Loa, Hawaii. The predictions of our model indicate the importance of abiotic conditions on mosquito dynamics and have important implications for the management of diseases transmitted by Cx. quinquefasciatus in Hawaii and elsewhere.

Culex quinquefasciatus Say (Diptera: Culicidae) is a widely distributed mosquito vector species in India and also in other tropical regions of the world. This species is implicated in the transmission of lymphatic filariasis in many countries. This species is reported to be widely resistant to insecticides of different classes in current use. In the present study, bio-efficacy of chlorfenapyr, an insecticide of pyrrole class with a novel mode of action was tested for the control of Cx. quinquefasciatus. Studies were performed to determine the diagnostic dosage; residual efficacy on different artificially fabricated substrates, namely wood, mud, mud+lime, cement and cement+distemper; to assess cross-resistance with different insecticides; and synergism/antagonism using piperonyl butoxide (PBO). A dosage of 5.0% chlorfenapyr was determined as diagnostic dosage with 2 h exposure and 48 h holding period for assessing the susceptibility of mosquitoes. The residual efficacy was observed up to 34 weeks on wood and mud+lime substrates while on other substrates, it was about 15 weeks at a dosage of 400mg a.i./m(2). Laboratory-reared strains of Cx. quinquefasciatus showed cross-resistance, whereas field-collected mosquitoes showed absence of cross-resistance to chlorfenapyr. Potentiation bioassays showed antagonistic effect of PBO to chlorfenapyr toxicity owing to the involvement of oxidases in the initial step of a conversion of pro-insecticide chlorfenapyr to toxic form CL 303268. The present study results have shown that chlorfenapyr can be a potential insecticide for the control of multiple insecticide resistant strains of Cx. quinquefasciatus. However, in countries where indoor residual spray (IRS) is not targeted for the control of this species, like in India, chlorfenapyr used in IRS for the control of malaria vectors in rural and peri-urban areas can additionally provide control of Cx. quinquefasciatus also.

The toxicities of 10 insecticides were examined against late third instar Culex tritaeniorhynchus Giles (Diptera: Culicidae) using the direct-contact mortality bioassay. Six geospatially-distant field mosquitoes were collected from Chuncheon-si, Hwaseong, Seosan. Jeonju, Daegu, and Busan in the Republic of Korea. Marked regional variations of insecticide susceptibility were observed. Field populations of Seosan, Jeonju, and Daegu from agricultural areas showed higher to extremely higher insecticide susceptibility to pyrethroids than those of Chuncheon-si, Hwaseong, and Busan strains from non-agricultural areas. Extremely high to low levels of susceptibility were measured: bifenthrin, susceptible ratio (SR) = 2.7–896.3; β-cyfluthrin, SR = 1.8–633.3; α-cypermethrin, SR = 1.2–1,051.9; deltamethrin, SR = 1.3–711.1; permethrin, SR = 1.5–1,053.4; etofenprox, SR = 2.2–29.3; chlorfenapyr, SR = 5.1–103.6; chlorpyrifos, SR = 2.3– 337.0; fenitrothion, SR = 2.0–142.3; and fenthion, SR = 1.4–186.2. Cx. tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to the agricultural insecticides, and that's why the mosquito species demonstrated high resistance to pyrethroids, which were used for a long time to control agricultural pests in the localities. These results indicate that careful selection and rotational use of these insecticides may result in continued satisfactory control against field populations of Japanese encephalitis vector mosquitoes. PMID:23879898

The dispersal patterns of mosquito vectors are important drivers of vector-borne infectious disease dynamics and understanding movement patterns is pivotal to devise successful intervention strategies. Here, we investigate the dispersal patterns of two globally important mosquito vectors, Aedes albopictus and Culex quinquefasciatus, by marking naturally-occurring larvae with stable isotopes (13C or 15N). Marked individuals were captured with 32 CDC light trap, 32 gravid trap, and 16 BG Sentinel at different locations within two-kilometer radii of six larval habitats enriched with either 13C or 15N. In total, 720 trap nights from July to August 2013 yielded a total of 32,140 Cx. quinquefasciatus and 7,722 Ae. albopictus. Overall, 69 marked female mosquitoes and 24 marked male mosquitoes were captured throughout the study period. The distance that Cx. quinquefasciatus females traveled differed for host-seeking and oviposition-seeking traps, with females seeking oviposition sites traveling further than those seeking hosts. Our analysis suggests that 41% of Cx. quinquefasciatus females that were host-seeking occurred 1–2 kilometer from their respective natal site, while 59% remained within a kilometer of their natal site. In contrast, 59% of Cx. quinquefasciatus females that were seeking oviposition sites occurred between 1–2 kilometer away from their larval habitat, while 15% occurred > 2 kilometer away from their natal site. Our analysis estimated that approximately 100% of Ae. albopictus females remained within 1 km of their respective natal site, with 79% occurring within 250m. In addition, we found that male Ae. albopictus dispersed farther than females, suggesting male-biased dispersal in this Ae. albopictus population. This study provides important insights on the dispersal patterns of two globally relevant vector species, and will be important in planning next generation vector control strategies that mitigate mosquito-borne disease through sterile insect

Control programs using conventional insecticides to target anthropogenic mosquito habitats are very expensive because these habitats are widespread, particularly in cities of most African countries. Additionally, there are serious environmental concerns regarding large-scale application of most conventional insecticides. Clearly there is a need for alternative methods that are more effective, less expensive, and environmentally friendly. One such method would be the application of preparations made from parts of the neem tree, Azadirachta indica A. Jussieu (Sapindales: Meliaceae). In this study, aqueous crude extracts and crude powder were prepared from different parts of neem, and the efficacies of the preparations on juvenile stages of Culex quinquefasciatus Say (Diptera: Culicidae) were evaluated in the laboratory. When larvae were exposed to a concentration of 0.1 g/mL extract for 24 hours, percent mean mortality (± SE) was 72.7 plusmn; 1.8 for the bark, 68.7 ± 1.6 for fruits and 60 ± 1.6 for leaves. These means were not significantly different (χ2 = 4.12; df = 2; p = 0.127). At a concentration of 0.01 g/mL, > 95% of the larvae died within 24 hours of exposure to powdered neem leaf, but it took 120 hours to reach the same level of larval mortality in aqueous leaf extract. The crude extract slowly inhibited the growth and development of mosquitoes while the crude powder acted more as a barrier; the mosquitoes probably died from suffocation. However, both types of preparations can be made and used by local people to control mosquito breeding in anthropogenic habitats, especially in urbanized areas. PMID:22233153

Organophosphate pesticides are widely used in vector mosquito management and agricultural pest management. These chemicals enter into natural water bodies and soil and cause hazards to the environment. The objective of this study was to prepare a natural pesticide which will not harm the environment and yet control vector mosquitoes. PONNEEM, a novel biopesticide, patented and prepared from the oils of Azadirachta indica and Pongamia glabra, was tested against Anopheles stephensi and Culex quinquefasciatus. One hundred percent larvicidal and ovicidal activities were observed at 0.1-ppm concentration of PONNEEM against the two mosquito species under laboratory and sunlight conditions up to 12 months from the date of manufacture. Very high oviposition reduction of 26.46 and 32.16 % is also recorded. Reductions in α-esterase level (0.0818 ± 0.340 and 0.2188 ± 0.003), β-esterase level (0.0866 ± 0.026 and 0.0398 ± 0.010 μg naphthol produced/min/mg larval protein), glutathione S-transferase enzyme (14.2571 ± 0.51 and 15.3326 ± 0.51 μmol/min/mg larval protein) and total protein levels (0.0390 ± 0.008 and 0.1975 ± 0.029 mg/individual larva in treated groups of A. stephensi and C. quinquefasciatus at 0.1-ppm concentration, respectively. The non-target organisms such as Gambusia affinis and Diplonychus indicus were not affected. Biopesticides are good alternatives to synthetic pesticides. PONNEEM can be effectively used for the management of human vector mosquitoes. Since it has a biodegradable nature and does not alter the environmental condition of water and soil.

Background & objectives: Mosquitoes transmit serious human diseases, causing millions of deaths every year and the development of resistance to chemical insecticides resulting in rebounding vectorial capacity. Plants may be alternative sources of mosquito control agents. The present study assessed the role of larvicidal activities of hexane, chloroform, ethyl acetate, acetone, and methanol dried leaf and bark extracts of Annona squamosa L., Chrysanthemum indicum L., and Tridax procumbens L. against the fourth instar larvae of malaria vector, Anopheles subpictus Grassi and Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae). Methods: Larvicidal activities of three medicinal plant extracts were studied in the range of 4.69 to 1000 mg/l in the laboratory bioassays against early 4th instar larvae of An. subpictus and Cx. tritaeniorhynchus. The mortality data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the respective species. Results: All plant extracts showed moderate effects after 24 h of exposure; however, the highest toxic effect of bark methanol extract of A. squamosa, leaf ethyl acetate extract of C. indicum and leaf acetone extract of T. procumbens against the larvae of An. subpictus (LC50 = 93.80, 39.98 and 51.57 mg/l) and bark methanol extract of A. squamosa, leaf methanol extract of C. indicum and leaf ethyl acetate extract of T. procumbens against the larvae of Cx. tritaeniorhynchus (LC50 =104.94, 42.29 and 69.16 mg/l) respectively. Interpretation & Conclusions: Our data suggest that the bark ethyl acetate and methanol extract of A. squamosa, leaf ethyl acetate and methanol extract of C. indicum, acetone and ethyl acetate extract of T. procumbens have the potential to be used as an ecofriendly approach for the control of the An. subpictus, and Cx. tritaeniorhynchus. PMID:21808141

Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikunguniya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The adulticidal and repellent activities of crude hexane, ethyl acetate, benzene, chloroform, and methanol extracts of leaf of Eclipta alba and Andrographis paniculata were assayed for their toxicity against two important vector mosquitoes, viz., Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae). The adult mortality was observed after 24 h of exposure. All extracts showed moderate adulticide effects; however, the highest adult mortality was found in methanol extract of A. paniculata against the adults of C. quinquefasciatus and A. aegypti with the LC(50) and LC(90) values were 149.81, 172.37 ppm and 288.12, 321.01 ppm, respectively. The results of the repellent activity of hexane, ethyl acetate, benzene, chloroform, and methanol extract of E. alba and A. paniculata plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm(2) were applied on skin of forearm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal ecofriendly approach for the control of mosquitoes. This is the first report on the mosquito adulticidal and repellent activities of the reported E. alba and A. paniculata plants.

Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 μg/mL), temephos (0.002 μg/mL) and permethrin (0.01 μg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure.

Culex tarsalis Coquillett females were infected with the NY99 strain of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) and then incubated under constant temperatures of 10-30 degrees C. At selected time intervals, transmission was attempted using an in vitro capillary tube assay. The median time from imbibing an infectious bloodmeal until infected females transmitted WNV (median extrinsic incubation period, EIP50) was estimated by probit analysis. By regressing the EIP rate (inverse of EIP50) as a function of temperature from 14 to 30 degrees C, the EIP was estimated to require 109 degree-days (DD) and the point of zero virus development (x-intercept) was estimated to be 14.3 degrees C. The resulting degree-day model showed that the NY99 WNV strain responded to temperature differently than a lineage II strain of WNV from South Africa and approximated our previous estimates for St. Louis encephalitis virus (family Flaviviridae, genus Flavivirus, SLEV). The invading NY99 WNV strain therefore required warm temperatures for efficient transmission. The time for completion of the EIP was estimated monthly from temperatures recorded at Coachella Valley, Los Angeles, and Kern County, California, during the 2004 epidemic year and related to the duration of the Cx. tarsalis gonotrophic cycle and measures of WNV activity. Enzootic WNV activity commenced after temperatures increased, the duration of the EIP decreased, and virus potentially was transmitted in two or less gonotrophic cycles. Temperatures in the United States during the epidemic summers of 2002-2004 indicated that WNV dispersal and resulting epicenters were linked closely to above-average summer temperatures.

Spinosad 12% suspension concentrate (SC) and 0.5% granular (G) formulations were tested against Culex quinquefasciatus immatures at 3 dosages--50, 100, and 150 mg active ingredient (AI)/m2--in cesspits, street drains, and disused wells in comparison with temephos 1% G at 1 part per million. The 2 formulations reduced the density of pupae of Cx. quinquefasciatus by 80-100% for 7-14 days in cesspits, 7-18 days in drains, and 33-69 days in disused wells at the 3 dosages tested. The 12% SC and 0.5% G formulations were equally effective in suppressing the larval and pupal populations of Cx. quinquefasciatus in all the habitats treated. At 100 mg AI/m2 in drains and cesspits, the efficacy of spinosad 0.5% G was greater than that of temephos (1% G) and that of spinosad 12% SC was comparable to that of temephos (1% G) applied to the 2 habitats. In disused wells, both the formulations lasted longer than temephos. Based on the small-scale trial, the rate of 150 mg AI/m2 of spinosad was selected for field application to cesspits and drains and 50 mg AI/m2 for application to disused wells. The efficacy observed in medium-scale trials was in agreement with that of small-scale trials. The trial showed that the 2 formulations of spinosad can be used for larval control of Cx. quinquefasciatus, at 50 mg AI/m2, applied at weekly intervals to cesspits and street drains and at monthly intervals to disused wells. At the higher dose of 150 mg AI/m2, the 2 formulations were effective in suppressing the pupal population for 2 wk in cesspits and street drains and for 2 months in disused wells.

Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 μg/mL), temephos (0.002 μg/mL) and permethrin (0.01 μg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure. PMID:28085898

Control programs using conventional insecticides to target anthropogenic mosquito habitats are very expensive because these habitats are widespread, particularly in cities of most African countries. Additionally, there are serious environmental concerns regarding large-scale application of most conventional insecticides. Clearly there is a need for alternative methods that are more effective, less expensive, and environmentally friendly. One such method would be the application of preparations made from parts of the neem tree, Azadirachta indica A. Jussieu (Sapindales: Meliaceae). In this study, aqueous crude extracts and crude powder were prepared from different parts of neem, and the efficacies of the preparations on juvenile stages of Culex quinquefasciatus Say (Diptera: Culicidae) were evaluated in the laboratory. When larvae were exposed to a concentration of 0.1 g/mL extract for 24 hours, percent mean mortality (± SE) was 72.7 plusmn; 1.8 for the bark, 68.7 ± 1.6 for fruits and 60 ± 1.6 for leaves. These means were not significantly different (χ(2) = 4.12; df = 2; p = 0.127). At a concentration of 0.01 g/mL, > 95% of the larvae died within 24 hours of exposure to powdered neem leaf, but it took 120 hours to reach the same level of larval mortality in aqueous leaf extract. The crude extract slowly inhibited the growth and development of mosquitoes while the crude powder acted more as a barrier; the mosquitoes probably died from suffocation. However, both types of preparations can be made and used by local people to control mosquito breeding in anthropogenic habitats, especially in urbanized areas.

Synthetic insecticides are generally employed to control the mosquito population. However, their injudicious over usage and non-biodegradability are associated with many adverse effects on the environment and mosquitoes. The application of environment-friendly mosquitocidals might be an alternate to overcome these issues. In this study, we found that organic or aqueous extracts of Agave angustifolia leaves exhibited a strong larvicidal activity (LD50 28.27 μg/ml) against Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi larvae within a short exposure of 12 h. The larvicidal activity of A. angustifolia is inherited and independent of the plants vegetative growth. Interestingly, the plant larvicidal activity was observed exclusively during the summer season (April–August, when outside temperature is between 30 and 50°C) and it was significantly reduced during winter season (December–February, when the outside temperature falls to ~4°C or lower). Thus, we hypothesized that the larvicidal components of A. angustifolia might be induced by the manipulation of environmental temperature and should be resistant to the hot conditions. We found that the larvicidal activity of A. angustifolia was induced when plants were maintained at 37°C in a semi-natural environment against the controls that were growing outside in cold weather. Pre-incubation of A. angustifolia extract at 100°C for 1 h killed 60% larvae in 12 h, which gradually increased to 100% mortality after 24 h. In addition, the dry powder formulation of A. angustifolia, also displayed a strong larvicidal activity after a long shelf life. Together, these findings revealed that A. angustifolia is an excellent source of temperature induced bioactive metabolites that may assist the preparedness for vector control programs competently. PMID:26793700

Introduced mosquito-borne pathogens avian malaria (Plasmodium relictum Grassi and Feletti) and avian pox virus (Avipoxvirus) have been implicated in the past extinctions and declines of Hawaiian avifauna and remain significant obstacles to the recovery and restoration of endemic Hawaiian birds. Effective management of avian disease will require extensive mosquito control efforts that are guided by the local ecology of the vector Culex quinquefasciatus Say (Diptera: Culicidae). During October and November 1997 and September through November 1998 five mark-release-recapture experiments with laboratory-reared Cx. quinquefasciatus were conducted in a native rain forest on Hawaii Island. Of the overall 66,047 fluorescent dye-marked and released females, 1,192 (1.8%) were recaptured in 43-52 CO2-baited traps operated for 10-12-d trapping periods. Recaptured mosquitoes were trapped in all directions and at distances up to 3 km from the release site. The cumulative mean distance traveled (MDTs) over the trapping period ranged from a high of 1.89 km after 11 d (September 1998) to a low of 0.81 km after 11 d (November 1998). Released mosquitoes moved predominately in a downwind direction and they seemed to use forestry roads as dispersal corridors. Applying an estimated MDT of 1.6 km to a geographical information system-generated map of the Hakalau Forest National Wildlife Refuge clearly demonstrated that the effective refuge area could be reduced 60% by mosquitoes infiltrating into managed refuge lands. These findings should have significant implications for the design of future refuges and development of effective mosquito-borne avian disease control strategies.

Culex quinquefasciatus Say is an important disease vector throughout much of the world. Experiments were conducted to determine the effects of different larval habitat substrates on the fitness and biting efficiency of Cx. quinquefasciatus adults. Our findings indicate that the development time (egg to adult) of larvae reared in irrigation water was 8.63 d while that of larvae reared in distilled water was 17.10 d (Effect size = 0.95). However, the rate of adult emergence was similar for all the tested treatments. Furthermore, the mean weight of an egg raft varied between larval habitats: distilled water (1.83 mg), rainfall water (1.25 mg), irrigation water (1.52 mg), and sewerage water (2.52 mg) (Effect size = 0.91). But, the fecundity (eggs per female) and hatchability (%) were statistically similar in all the rearing mediums (Effect size = 0.79). Longevity of females in all the tested populations did not differ significantly (Effect size = 0.91). The mean relative growth rates of larvae reared in tap water (0.80) and distilled water (0.86) habitats were lower than growth rates in all other rearing habitats (Effect size = 0.96). The intrinsic rate of natural increase in tap water (0.27) and irrigation water (0.35) was significantly higher than that in distilled water (0.09) and sewerage water (0.16) (Effect size = 0.84). Adults reared in rain water had the highest biting efficiency among all the tested populations. These results provide useful information for the management of Cx. quinquefasciatus.

Evoked potential measures are integral to the treatment of patients with cochlear implants. In particular, these techniques are useful in the management of the pediatric patient. This brief report describes three categories of evoked potentials including clinical and research examples: electrically evoked auditory brain stem responses with transtympanic stimulation, middle-latency responses with cochlear implant stimulation, and cognitive evoked potentials elicited by speech stimuli.